Peptides and compounds that bind selectins including endothelial leukocyte adhesion molecule 1

ABSTRACT

Disclosed are peptides and peptide mimetics that bind selectins, including endothelial leukocyte adhesion molecule 1 (ELAM-1). Such peptides and peptide mimetics are useful in methods for blocking adhesion of leukocytes to the selectins for the purpose of inhibiting inflammation as well as in diagnostic methods employing labeled peptides and peptide mimetics that bind selectins for the purpose of determining the site of inflammation in mammals which inflammation is mediated by the presence of one or more selectins.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation-in-part of U.S. application Ser. No.08/057,295 filed May 5, 1993, now abandoned which, in turn, is acontinuation-in-part of U.S. application Ser. No. 07/881,395 filed May6, 1992 now abandoned, the disclosures of both of which are incorporatedherein by reference in their entirety.

BACKGROUND OF THE INVENTION

1. Field of the Invention

This invention is directed to peptides and peptide mimetics that bindselectins, including endothelial leukocyte adhesion molecule 1 (ELAM-1).This invention is further directed to methods for blocking adhesion ofleukocytes to the selectins for the purpose of inhibiting inflammation,inhibiting tumor metastasis, inhibiting reperfusion injury, and thelike. This invention is still further directed to diagnostic methodsemploying labeled peptides and peptide mimetics that bind selectins forthe purpose of determining the site of inflammation in mammals whichinflammation is mediated by the presence of one or more selectins.

References

The following references are cited in this application as superscriptnumbers at the relevant portion of the application:

1 Bevilacqua, et al., Proc. Natl. Acad. Sci. USA, 84:9238-9242 (1987)

2 Geng, et al., Nature, 343:757-760 (1990)

3 Springer, Nature, 346:425-434 (1990)

4 Bevilacqua, et al., Cell, 67:233 (1991)

5 Johnston, et al., Cell, 56:1033-1044 (1989)

6 Paulson, Selectin/Carbohydrate-Mediated Adhesion of Leukocytes inAdhesion: Its Role in Inflammatory Disease, pp. 19-42

7 Springer, et al., Nature, 349:196-197 (1991)

8 Lowe, et al., Cell, 63:475-485 (1990)

9 Phillips, et al., Science, Vol. 250:1130-1132 (1990)

10 Walz, et al., Science, 250:1132 et seq. (1990)

11 Larsen, et al., Cell, 63:467-474 (1990)

12 Brandley, et al., PCT International Patent Application No. WO92/02527, published Feb. 20, 1992

13 Furie, et al., PCT International Patent Application No. WO 92/16612,published Oct. 1, 1992

14 Phillips, et al, Science, 250:1130-1135 (1990)

15 Parekh, Oxford GlycoSystems Ltd., Tech. Bull. 11, (1991)

16 Tiemeyer, et al., Proc. Natl. Acad. Sci. USA, 85.:1138-1142 (1991)

17 Tyrrell, et al., Proc. Natl. Acad. Sci. USA, 88:10372-10376 (1991)

18 Polley, et al., Proc. Natl. Acad. Sci. USA, 88:6224-6228 (1991)

19 Paulson, et al., PCT International Patent Application Publication No.WO 91/19502, published Dec. 26, 1991

20 McEver, PCT International Patent Application Publication No. WO92/01718, published Feb. 6, 1992

21 Edgington, Biotechnology, 10:383-389 (1992)

22 McEver, U.S. Pat. No. 5,198,424 for Functionally ActiveSelectin-Derived Peptides, issued Mar. 30, 1993

23 Heavner, et al., PCT International Patent Application Publication No.WO 93/05070 published Mar. 18, 1993

24 Brandley, et al., U.S. Pat. No. 5,143,712, for Method for Determininga Cite of Inflammation Using ELAM-1 Ligands, issued Sep. 1, 1992

25 Bender and Lee, Ann. Rep. Med. Chem., 25:185-193 (1989)

26 Hannum, et al., U.S. Pat. No. 5,075,222, for Interleukin-1Inhibitors, issued Dec. 24, 1991

27 Cwirla, et al., Proc. Natl. Acad. Sci. USA, 87:6378-6382 (1990)

28 Pirrung, et al., U.S. Pat. No. 5,143,854, for Large ScalePhotolithographic Solid Phase Synthesis of Polypeptides and ReceptorBinding Screening Thereof, issued Sep. 1, 1992

29 Pirrung, et al., PCT International Patent Application Publication No.90/15070, published Dec. 13, 1990

30 Fodor, et al., Science, 251:767-773 (1991)

32 Dower, et al., Ann. Rep. Med. Chem., 26:271-280 (1991)

33 Caras and Weddell, Science, 243:1196-1198, (1989)

34 Lin, et al., Science, 249:677-679 (1990)

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38 Gisin, Helv. Chim. Acta, 56:1487 (1973)

39 Cho, et al., Science, 261:1303-1305 (1993)

40 Stewart, Solid Phase Peptide Syntheses (Freeman and Co., SanFrancisco, 1969).

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The disclosure of all publications, patents and patent applicationscited in this application are herein incorporated by reference in theirentirety to the same extent as if each individual publication or patentapplication was specifically and individually indicated to beincorporated by reference in its entirety.

State of the Art

The accumulation of blood leukocytes at sites of mammalian inflammationdepends upon the localization of these leukocytes by adhesion to thevascular endothelium and subsequent extravasation of the leukocytes.Certain cytokines, such as interleukin 1 (IL-1) and tumor necrosisfactor alpha and beta (TNF-α and TNF-β respectively), as well asbacterial endotoxin, have been shown to act on cultured humanendothelial cells to increase leukocyte adhesion. Bevilacqua, et al.¹report the identification of an inducible endothelial cell surfaceprotein with a molecular weight of about 115 kD, designated endothelialleukocyte adhesion molecule 1 (ELAM-1, also known as LECAM-2, CD62E, andE-selectin) which protein is involved in the process of leukocyteadhesion to the vascular endothelium prior to extravasation ofleucocytes through the endothelium to the site of inflammation.

Further studies have shown that the E-selectin is a member of a familyof receptors referred to as "selectins" which family includes otherknown receptors such as PADGEM (also known as the P-selectin, GMP-140,LECAM-3 and CD62P) and LAM-1 (also known as the L-selectin, LECAM-1,MEL-14 and CD62L)²,3,4,5. Each of these other selectins is also involvedin the process of leukocyte adhesion to the vascular endothelium priorto extravasation of leukocytes through the endothelium to the site ofinflammation.

Homologous to each of these known selectin proteins is an N-terminallectin domain, an epidermal growth factor (EGF) like domain, a number ofrepeating units related to those in complement binding proteins, atransmembrane domain, and a short cytoplasmic tail⁶. Evidence suggeststhat the in vivo ligands for these selectin proteins are glycoconjugateswhich purportedly bind to the selectin through, at least in part, thecarbohydrate portion of the glycoconjugate. The carbohydrate structuresof such ligands include structures containing terminally linkedαNeu5Ac(2→3)βGal(1→4)-[α-L-Fuc (1→3)]-βGlcNAc- (sialyl Lewis^(x) orSLe^(x)) which structures are part of the cell surface glycoprotein andglycolipid groups of neutrophils for the E- and the P-selectin and ofendothelial cells for the L-selectin during adhesion.⁷⁻¹⁸

In view of their binding to selectins, the administration of SLe^(x) andrelated carbohydrates has been proposed as a means for controllinginflammatory conditions and other conditions mediated by selectinproteins by interfering with leukocyte adhesion to theseproteins¹²⁻¹³,19-20. Such inflammatory conditions mediated by selectinproteins include, by way of example, (1) dermal inflammation, includingIl-1-induced skin inflammation; atopic dermatitis, psoriasis, andallergic contact dermatitis; delayed-type hypersensitivity reactioninduced by DNCB on human skin; and endotoxin-induced acute cutaneousinflammation in baboons; (2) intestinal inflammation, including inflamedcolonic mucosa of patients with ulcerative or Crohn's colitis;inflammatory bowel disease; and intestinal graft-versus-host disease;(3) other sites of inflammation, including synovial tissues frompatients with rheumatoid arthritis and osteo arthritis; ratendotoxin-induced uveitis, and inflamed endothelium of asthma model(primate antigen inhalation), acute respiratory distress syndome (ARDS);and (4) septic shock.²⁵,26

Central to such disclosures is the administration to the patient of acarbohydrate structure which binds to the selectin thereby preventingbinding of the selectin protein with the carbohydrate binding structuresfound on the cell surface of the neutrophils or on the endothelium.

It has also been disclosed to use such carbohydrates for diagnosticpurposes in identifying in vivo the sites of the E-selectinexpression²³,24. Such information is important in the treatment ofE-selectin mediated conditions such as tissue damage associated withmyocardial infarction, inflammation, etc. Additionally, it has also beensuggested to couple anti-inflammatory drugs to the carbohydrate ligandsof E-selectin to provide for site specific drug delivery to the point ofinflammation¹⁹,24.

The use of carbohydrate structures as part of either the treatmentregimen for E-selectin mediated conditions (e.g., inflammatoryconditions) or diagnosis of sites of E-selectin mediated conditions iscomplicated, however, by the fact that these compounds are difficult tosynthesize and, using conventional synthetic procedures, costs forpreparation of such carbohydrates are prohibitive²¹.

Contrarily, it has also been suggested to use polyclonal/monoclonalantibodies to selectins, e.g., E-selectin, as well as peptides derivedfrom the lectin-like domain of the P-selectin for the purpose ofinterfering with leukocyte/selectin binding²²,23 in a therapeuticregimen.

In the case of peptides derived from the lectin-like domain of theP-selectin, it is disclosed that these materials have the bindingproperties of selectins and can bind neutrophils. When administered,these peptides compete with the selectins for binding to neutrophils andsuch competition can be therapeutically used to inhibit conditionsmediated by selectin/neutrophil interactions such as, for example,inflammatory conditions.

In the case of polyclonal/monoclonal antibodies, it has been disclosedthat these materials bind directly to the selectin thereby inhibitingthe ability of neutrophils to bind to selectins. Inhibition of theneutrophil/selectin binding can be used therapeutically for conditionsmediated by selectins.

However, therapeutic use of such polyclonal/monoclonal antibodies is notfavored because such antibodies are typically immunogenic andadministration is usually limited to a single occurrence²². In suchcases, additional administrations of such antibodies can produce anadverse reaction in the patient.

Likewise, the use of peptides having binding properties of selectins isnot favored because such peptides typically do not bind selectins and,accordingly, are not suitable for use in the in vivo diagnosis of sitesof E-selectin expression which correlates to disease conditions mediatedby E-selectin including inflammatory conditions. Nor can these peptidesbe used for the site specific delivery of anti-inflammatory or otherdrugs suitable for treatment of conditions mediated by E-selectin.

SUMMARY OF THE INVENTION

This invention is directed, in part, to the novel and unexpecteddiscovery that defined low molecular weight peptides and peptidemimetics have strong binding properties to E-selectin and, in somecases, to the other selectins, i.e., the P-selectin and L-selectin.Accordingly, such peptides and peptide mimetics are useful fortherapeutic purposes in treating conditions mediated by E-selectin(e.g., inflammatory conditions) as well as for diagnostic purposes inidentifying in vivo the vascular site of E-selectin protein and, hence,the site of conditions mediated by E-selectin expression. Such peptidesand peptide mimetics can also be coupled to anti-inflammatory drugs orother drugs suitable for treatment of the disease conditions mediated byE-selectin expression so as to provide for in vivo site specific drugdelivery to the point of E-selectin expression.

Peptides and peptide mimetics suitable for diagnostic purposes areCharacterized by having a molecular weight of less than about 2000daltons and a binding affinity to E-selectin as expressed by an IC₅₀-HL60 of about no more than about 100 μM as determined by the HL60binding affinity assay set forth in Example 1 and preferably no morethan 50 μM.

In one preferred embodiment, the molecular weight of the peptide orpeptide mimetic is from about 250 to about 1800 daltons.

In another preferred embodiment, the IC₅₀ binding affinity of thepeptide or peptide mimetic is determined by binding competition betweena candidate peptide and a standard as per the in vitro procedureprovided in Example 2 hereinbelow. This example measures theconcentration of peptide or peptide mimetic required to inhibit 50% ofthe binding of a radiolabeled standard to E-selectin and the resultingIC₅₀ value is referred to as the "IC₅₀ -standard". In either case, alower IC₅₀ -HL60 or a lower IC₅₀ -standard for the peptide or peptidemimetic correlates to a stronger binding affinity to E-selectin.

Preferably, the peptide or peptide mimetic has an IC₅₀ -standard ofabout 2.5 mM or less. For therapeutic purposes, these peptides andpeptide mimetics preferably have a molecular weight of less than about2000 daltons and a binding affinity to E-selectin as expressed by anIC₅₀ -standard of about 2.0 mM or less. In a more preferred embodiment,the peptide or peptide mimetic has an IC₅₀ -standard of about 2.5 μM orless and even more preferably has an IC₅₀ -standard of about 10 nM orless.

When used for diagnostic purposes, the peptides and peptide mimeticspreferably are labeled with a detectable label and, accordingly, thepeptides and peptide mimetics without such a label serve asintermediates in the preparation of labeled peptides and peptidemimetics.

Peptides meeting the defined criteria for molecular weight and bindingaffinity for E-selectin comprise 3 or more amino acids wherein the aminoacids are naturally occurring or synthetic (non-naturally occurring)amino acids. Peptide mimetics include peptides having one or more of thefollowing modifications:

peptides wherein one or more of the peptidyl [C(O)NH--] linkages (bonds)have been replaced by a non-peptidyl linkage such as a --CH₂ -carbamatelinkage [--CH₂ --OC(O)NR--], a phosphonate linkage, a --CH₂ -sulfonamide[--CH₂ --S(O)₂ NR--] linkage, a urea [--NHC(O)NH--] linkage, a --CH₂-secondary amine linkage, or with an alkylated peptidyl linkage[--C(O)NR⁶ -- where R⁶ is lower alkyl],

peptides wherein the N-terminus is derivatized to a --NRR¹ group, to a--NRC(O)R group, to a --NRC(O)OR group, to a --NRS(O)₂ R group, to a--NHC(O)NHR group where R and R¹ are hydrogen or lower alkyl with theproviso that R and R¹ are not both hydrogen, to a succinimide group,i.e., ##STR1## to a benzyloxycarbonyl-NH-- (CBZ--NH--) group, i.e.,##STR2## to a benzyloxycarbonyl-NH-- group having from 1 to 3substituents on the phenyl ring selected from the group consisting oflower alkyl, lower alkoxy, chloro, and bromo,

peptides wherein the C terminus is derivatized to --C(O)R² where R² isselected from the group consisting of lower alkoxy, and --NR³ R⁴ whereR³ and R⁴ are independently selected from the group consisting ofhydrogen and lower alkyl.

Accordingly, in a first compositional aspect, this invention is directedto peptides and peptide mimetics comprising:

a molecular weight of less than about 2000 daltons, and

a binding affinity to endothelial leukocyte adhesion molecule 1 asexpressed by an IC₅₀ -HL60 of no more than about 100 μM

wherein from zero to all of the --C(O)NH-- linkages of the peptide havebeen replaced by a linkage selected from the group consisting of a --CH₂OC(O)NR-- linkage, a phosphonate linkage, a --CH₂ S(O)₂ NR-- linkage, a--CH₂ NR-- linkage, a --C(O)NR⁶ -- linkage, and a --NHC(O)NH-- linkagewhere R is hydrogen or lower alkyl, and R⁶ is lower alkyl,

further wherein the N-terminus of said peptide or peptide mimetic isselected from the group consisting of a --NRR¹ group, a --NRC(O)R group,a --NRC(O)OR group, a --NRS(O)₂ Rgroup, a --NHC(O)NHR group, asuccinimide group, a benzyloxycarbonyl-NH-- group, and abenzyloxycarbonyl-NH-- group having from 1 to 3 substituents on thephenyl ring selected from the group consisting of lower alkyl, loweralkoxy, chloro, and bromo, where R and R¹ are independently selectedfrom the group consisting of hydrogen and lower alkyl,

and still further wherein the C-terminus of said peptide or peptidemimetic has the formula --C(O)R² where R² is selected from the groupconsisting of hydroxy, lower alkoxy, and --NR³ R⁴ where R³ and R⁴ areindependently selected from the group consisting of hydrogen and loweralkyl and where the nitrogen atom of the --NR³ R⁴ group can optionallybe the amine group of the N-terminus of the peptide so as to form acyclic peptide

and physiologically acceptable salts thereof.

In a second compositional aspect, this invention is directed to alabeled peptide or peptide mimetic comprising:

a molecular weight of less than about 2000 daltons,

a binding affinity to endothelial leukocyte adhesion molecule 1 asexpressed by an IC₅₀ -HL60 of about 100 μM or less, and

having covalently attached thereto a label capable of detection

wherein from zero to all of the --C(O)NH-- linkages of the peptide havebeen replaced by a linkage selected from the group consisting of a --CH₂OC(O)NR-- linkage, a phosphonate linkage, a --CH₂ S(O)₂ NR-- linkage, a--CH₂ NR-- linkage, a --C(O)NR⁶ -- linkage, and a --NHC(O)NH-- linkagewhere R is hydrogen or lower alkyl, and R⁶ is lower alkyl,

further wherein the N-terminus of said peptide or peptide mimetic isselected from the group consisting of a --NRR¹ group, a --NRC(O)R group,a --NRC(O)OR group, a --NRS(O)₂ R group, a --NHC(O)NHR group, asuccinimide group, a benzyloxycarbonyl-NH-- group, and abenzyloxycarbonyl-NH-- group having from 1 to 3 substituents on thephenyl ring selected from the group consisting of lower alkyl, loweralkoxy, chloro, and bromo, where R and R¹ are independently selectedfrom the group consisting of hydrogen and lower alkyl,

and still further wherein the C-terminus of said peptide or peptidemimetic has the formula --C(O)R² where R² is selected from the groupconsisting of hydroxy, lower alkoxy, and --NR³ R⁴ where R³ and R⁴ areindependently selected from the group consisting of hydrogen and loweralkyl and where the nitrogen atom of the --NR³ R⁴ group can optionallybe the amine group of the N-terminus of the peptide so as to form acyclic peptide

and physiologically acceptable salts thereof.

Preferably, the peptides and peptide mimetics described above have anIC₅₀ -standard, as measured in Example 2 hereinbelow, of about 2.5 mM orless. Peptides having such an IC₅₀ -standard define a subclass ofpeptides meeting the IC₅₀ -HL60 requirements of 100 μM or less whichhave stronger binding affinity to endothelial leukocyte adhesionmolecule 1. In a more preferred embodiment, the peptide or peptidemimetic has an IC₅₀ -standard of about 2.0 μM or less and even morepreferably has an IC₅₀ -standard of about 10 nM or less.

Preferred peptides for use in this invention include peptides having acore structure comprising:

WXXLWXXM (SEQ ID NO 1)

where each amino acid is indicated by the one letter amino acid symbolsrecommended by the IUPAC-IUB Biochemcial Nomenclature Commission where Wis tryptophan, L is leucine, M is methionine, and X is any amino acid.

Other preferred peptides include peptides 9 to 12 amino acid residues inlength and comprising the sequence X₁ X₂ X₃ WX₄ X₅ LWX₆ X₇ X₈ X₉ (SEQ IDNO 2), wherein each residue can be independently selected as follows:X₁, is H, E, or D; X₂ is I, M, or Nle; X₃ is T or S; X₄ is D, A, E, orL; X₅ is Q or E; X₆ is N or D; X₇ is L, M, V, or I; X₈ is M or Nle; andX₉ is N, S, K, or Q. X₈ can also include methionine derivatives such asO-methyl methionine (MOCH₃) and met-sulfone (MSO₂ CH₃) as describedbelow.

Still other preferred peptides of the invention include peptides thatcomprise the following sequences:

DITWDQLWDLMK (SEQ ID NO 3)

DGDITWDQLWDLMK (SEQ ID NO 4)

DYTWFELWDMMQ (SEQ ID NO 5)

DITWDELWKIMN (SEQ ID NO 6)

QITWAQLWNMMK (SEQ ID NO 7)

DYSWHDLWEMMS (SEQ ID NO 8)

DITWDQLWDLNleK (SEQ ID NO 9)

HITWDQLWRIMT (SEQ ID NO 10)

d-DITWDQLWDLMK

Dd-ITWDQLWDLMK

ITWDQLWDLMK (SEQ ID NO 160)

HITWDQLWNVMN (SEQ ID NO 23)

wherein d- indicates a D-amino acid.

Other peptides suitable for use in this invention include peptides whichcomprise the following sequences:

YDDVCCELLF (SEQ ID NO 11)

DLPQWYTEWC (SEQ ID NO 12)

ENSHWCTCPC (SEQ ID NO 13)

DIEQDWVTWM (SEQ ID NO 14)

NEWCVVPCRL (SEQ ID NO 15)

DIWQDWVRWM (SEQ ID NO 16)

DLWQDWVTWM (SEQ ID NO 17)

DLWQDWVHWM (SEQ ID NO 18)

DIWQDWVTWM (SEQ ID NO 19)

DIWQDWVKWM (SEQ ID NO 20)

DIWQDWVRWM-NH ₂ (SEQ ID NO 164)

DIWEDWVRWM (SEQ ID NO 21)

DIWQDWITWM (SEQ ID NO 22)

This invention also provides for pharmaceutical compositions comprisingpeptides and peptide mimetics and a pharmaceutically acceptable carrierwherein said peptides and peptide mimetics comprise:

a molecular weight of less than about 2000 daltons, and

a binding affinity to endothelial leukocyte adhesion molecule 1 asexpressed by an IC₅₀ -HL60 of no more than 100 μM

wherein from zero to all of the --C(O)NH-- linkages of the peptide havebeen replaced by a linkage selected from the group consisting of a --CH₂OC(O)NR-- linkage, a phosphonate linkage, a --CH₂ S(O)₂ NR-- linkage, a--CH₂ NR-- linkage, a --C(O)NR⁶ -- linkage, and a --NHC(O)NH-- linkagewhere R is hydrogen or lower alkyl, and R⁶ is lower alkyl,

further wherein the N-terminus of said peptide or peptide mimetic isselected from the group consisting of a --NRR¹ group, a --NRC(O)R group,a --NRC(O)OR group, a --NRS(O)₂ R group, a --NHC(O)NHR group, asuccinimide group, a benzyloxycarbonyl-NH-- group, and abenzyloxycarbonyl-NH-- group having from 1 to 3 substituents on thephenyl ring selected from the group consisting of lower alkyl, loweralkoxy, chloro, and bromo, where R and R¹ are independently selectedfrom the group consisting of hydrogen and lower alkyl,

and still further wherein the C-terminus of said peptide or peptidemimetic has the formula --C(O)R² where R² is selected from the groupconsisting of hydroxy, lower alkoxy, and --NR³ R⁴ where R³ and R⁴ areindependently selected from the group consisting of hydrogen and loweralkyl and where the nitrogen atom of the --NR³ R⁴ group can optionallybe the amine group of the N-terminus of the peptide so as to form acyclic peptide

and physiologically acceptable salts thereof.

This invention still further comprises methods for inhibiting thebinding of ligands to selectin receptors, including the binding ofSLe^(x) present on leukocytes to E-selectin, and methods for treatingdisease and disease symptoms which are mediated by the presence ofselectins which methods comprise the administration to a patient of asufficient amount of a pharmaceutical composition as described above soas to provide for from about 0.001 to about 20 milligrams of suchpeptides or peptide mimetics per kilogram of body weight daily in thepatient. Preferably, administration to the patient provides from about0.001 to about 15 milligrams and more preferably from about 0.001 toabout 10 milligrams of such peptide per kilogram of bodyweight daily.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 illustrates the results of a cell adhesion assay in the presenceof various peptides; the assay is described in Example 1.

FIG. 2A illustrates the results of a cell adhesion assay in the presenceof various concentrations of an E-selectin blocking peptide of theinvention; the assay is described in Example 1 and FIG. 2B illustratesthe results of the same assay in varying concentrations of a monofucosylmonosialyllactose.

FIGS. 3A and 3B illustrate two peptides on plasmids assay formats. InFIG. 3A, a binding assay is conducted in which the specificity of apeptide for immobilized E-selectin receptor is related to the amount ofsignal generated by alkaline phosphatase (AP) conjugated to anti-LacIantibodies in turn bound to LacI-peptide fusion proteins. In FIG. 3B,the IC₅₀ -standard of free peptides in solution is measured in acompetition assay.

FIG. 4 illustrates the results of a cell adhesion assay in the presenceof a preferred peptide of the invention, DITWDQLWDLMK (SEQ ID NO 3).

FIG. 5 illustrates that certain peptides having high binding specificityfor E-selectin can also bind other selectins such as P-selectin andL-selectin.

DETAILED DESCRIPTION OF THE INVENTION

This invention is directed to the discovery that certain defined lowmolecular weight peptides and peptide mimetics have strong bindingproperties to E-selectin and, accordingly, are useful in the treatmentof disease conditions mediated by E-selectin as well as in diagnosticassays wherein the presence of E-selectin is diagnostic for the site ofinflammation and/or other disease conditions mediated by E-selectinexpression.

However, prior to discussing this invention in further detail, thefollowing terms will first be defined.

Definitions

As used herein, the following terms have the following meanings:

The term "physiologically acceptable salts" refers to the non-toxicalkali metal, alkaline earth metal, and ammonium salts commonly used inthe pharmaceutical industry including the sodium, potassium, lithium,calcium, magnesium, barium, ammonium, and protamine zinc salts, whichare prepared by methods well known in the art. The term also includesnon-toxic acid addition salts, which are generally prepared by reactingthe compounds of this invention with a suitable organic or inorganicacid. Representative salts include the hydrochloride, hydrobromide,sulfate, bisulfate, acetate, oxalate, valerate, oleate, laurate, borate,benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate,succinate, tartrate, and napsylate salts, and the like. The particularsalt employed is not critical.

The term "lower alkyl" refers to straight and branched chain alkylgroups having from 1 to 6 carbon atoms, such as methyl, ethyl, n-propyl,iso-propyl, n-butyl, tert-butyl, sec-butyl, n-pentyl, n-hexyl,2-methylpentyl, and the like. Preferably, the lower alkyl group ismethyl or ethyl.

The term "lower alkoxy" refers to straight and branched chain alkoxygroups having from 1 to 6 carbon atoms, such as methoxy, ethoxy,n-propoxy, iso-propoxy, n-butoxy, tert-butoxy, sec-butoxy, n-pentoxy,n-hexoxy, 2-methylpentoxy, and the like. Usually, the lower alkoxy groupis methoxy or ethoxy.

Naturally occurring amino acid residues in peptides are abbreviated asrecommended by the IUPAC --IUB Biochemical Nomenclature Commission asfollows: Phenylalanine is Phe or F; Leucine is Leu or L; Isoleucine isIle or I; Methionine is Met or M; Norleucine is Nle; Valine is Val or V;Serine is Ser or S; Proline is Pro or P; Threonine is Thr or T; Alanineis Ala or A; Tyrosine is Tyr or Y; Histidine is His or H; Glutamine isGln or Q; Asparagine is Asn or N; Lysine is LyS or K; Aspartic Acid isAsp or D; Glutamic Acid is Glu or E; Cysteine is Cys or C; Tryptophan isTrp or W; Arginine is Arg or R; Glycine is Gly or G, and X is any aminoacid. Other naturally occurring amino acids include, by way of example,4-hydroxyproline, 5-hydroxylysine, and the like.

Synthetic or non-naturally occuring amino acids refer to amino acidswhich do not naturally occur in vivo but which, nevertheless, can beincorporated into the peptide structures described herein.

Preferred synthetic amino acids are the D-α-amino acids of naturallyoccurring L-α-amino acid as well as non-naturally occurring D- andL-α-amino acids represented by the formula H₂ NCHR⁵ COOH where R⁵ is 1)a lower alkyl group, 2) a cycloalkyl group of from 3 to 7 carbon atoms,3) a heterocycle of from 3 to 7 carbon atoms and 1 to 2 heteroatomsselected from the group consisting of oxygen, sulfur, and nitrogen, 4)an aromatic residue of from 6 to 10 carbon atoms optionally having from1 to 3 substituents on the aromatic nucleus selected from the groupconsisting of hydroxyl, lower alkoxy, amino, and carboxyl, 5)-alkylene-Y where alkylene is an alkylene group of from 1 to 7 carbonatoms and Y is selected from the group consisting of (a) hydroxy, (b)amino, (c) cycloalkyl and cycloalkenyl of from 3 to 7 carbon atoms, (d)aryl of from 6 to 10 carbon atoms optionally having from 1 to 3substituents on the aromatic nucleus selected from the group consistingof hydroxyl, lower alkoxy, amino and carboxyl, (e) heterocyclic of from3 to 7 carbon atoms and 1 to 2 heteroatoms selected from the groupconsisting of oxygen, sulfur, and nitrogen, (f) --C(O)R² where R² isselected from the group consisting of hydrogen, hydroxy, lower alkyl,lower alkoxy, and --NR³ R⁴ where R³ and R⁴ are independently selectedfrom the group consisting of hydrogen and lower alkyl, (g) --S(O)_(n) R⁶where n is an integer from 1 to 2 and R⁶ is lower alkyl and with theproviso that R⁵ does not define a side chain of a naturally occurringamino acid.

Other preferred synthetic amino acids include amino acids wherein theamino group is separated from the carboxyl group by more than one carbonatom such as β-alanine, γ-aminobutyric acid, and the like.

Particularly preferred synthetic amino acids include, by way of example,the D-amino acids of naturally occurring L-amino acids,L-1-napthyl-alanine, L-2-naphthylalanine, L-cyclohexylalanine, L-2-aminoisobutyric acid, the sulfoxide and sulfone derivatives of methionine##STR3## where n and R₆ are as defined above as well as the lower alkoxyderivative of methionine ##STR4##

The term "detectable label" refers to materials, which when covalentlyattached to the peptides and peptide mimetics of this invention, permitdetection of the peptide and peptide mimetics in vivo in the patient towhom the peptide or peptide mimetic has been administered. Suitabledetectable labels are well known in the art and include, by way ofexample, radioisotopes, fluorescent labels (e.g., fluorescein), and thelike. The particular detectable label employed is not critical and isselected relative to the amount of label to be employed as well as thetoxicity of the label at the amount of label employed. Selection of thelabel relative to such factors is well within the skill of the art.

Covalent attachment of the detectable label to the peptide or peptidemimetic is accomplished by conventional methods well known in the art.For example, when the ¹²⁵ I radioisotope is employed as the detectablelabel, covalent attachment of ¹²⁵ I to the peptide or the peptidemimetic can be achieved by incorporating the amino acid tyrosine intothe peptide or peptide mimetic and then iodating the peptide. Iftyrosine is not present in the peptide or peptide mimetic, incorporationof tyrosine to the N or C terminus of the peptide or peptide mimetic canbe achieved by well known chemistry. Likewise, ³² P can be incorporatedonto the peptide or peptide mimetic as a phosphate moiety through, forexample, a hydroxyl group on the peptide or peptide mimetic usingconventional chemistry.

Methodology

This invention provides peptides and peptide mimetics having strongbinding properties to E-selectin characterized as having a bindingaffinity to E-selectin as expressed by an IC₅₀ -HL60 of about 100 μM orless and more preferably by an IC₅₀ -standard of about 2.5 mM. Themethods described hereinbelow illustrate in vitro procedures foridentifying peptides having such binding properties and procedures foraltering such peptides to provide for peptide mimetics having suchbinding properties to E-selectin.

A. Identification of Peptides Which Bind E-Selectin

Peptides having a binding affinity to E-selectin can be readilyidentified by random peptide diversity generating systems coupled withan affinity enrichment process.

Specifically, random peptide diversity generating systems include boththe "peptides on plasmids" described in U.S. patent application Ser. No.07/963,321, filed on Oct. 15, 1992, which is a continuation-in-part ofU.S. patent application Ser. No. 07/778,233, filed on Oct. 16, 1991,(now U.S. Pat. No. 5,270,170) and the "peptides on phage" systemdescribed in U.S. patent application Ser. No. 07/718,577, filed Jun. 20,1991; in Cwirla et al.²⁷, in the "encoded synthetic library" (ESL)system described in U.S. patent application Ser. No. 07/946,239, filedSep. 18, 1992; and the "very large scale immobilized polymer synthesis"system described in U.S. patent application Ser. No. 5,143,85428;International Patent Application Serial No. 90/1507029; U.S. patentapplication Ser. No. 07/624,120, filed Dec. 6, 1990, Fodor et al.³⁰,Dower et al.³¹, and U.S. patent application Ser. No. 07/805,727,.filedDec. 6, 1991, the disclosures of each of which are incorporated hereinby reference in their entirety.

Using the procedures described above, random peptides were generallydesigned to have a defined number of amino acid residues in length(e.g., 12). To generate the collection of oligonucleotides encoding therandom peptides, the codon motif (NNK)_(x), where N is nucleotide A, C,G, or T (equimolar; depending on the methodology, other nucleotides canbe employed), K is G or T (equimolar), and x is an integer correspondingto the number of amino acids in the peptide (e.g., 12), was used tospecify any one of the 32 possible codons resulting from the NNK motif:1 for each of 12 amino acids, 2 for each of 5 amino acids, 3 for each of3 amino acids, and only one of the three stop codons. Thus, the NNKmotif encodes all of the amino acids, encodes only one stop codon, andreduces codon bias.

In the systems employed, the random peptides were presented either onthe surface of a phage particle, as part of a fusion protein comprisingeither the pIII or the pVIII coat protein of a phage fd derivative(peptides on phage), or as a fusion protein with the LacI peptide fusionprotein bound to a plasmid (peptides on plasmids).

The phage or plasmids, including the DNA encoding the peptides, wereidentified and isolated by an affinity enrichment process usingimmobilized E-selectin. The affinity enrichment process, sometimescalled "panning," involves multiple rounds of incubating the phage orplasmids with immobilized E-selectin, collecting the phage or plasmidsthat bind to E-selectin (along with the accompanying DNA), and producingmore of the phage or plasmids (along with accompanying LacI-peptidefusion protein) collected.

After several rounds of affinity enrichment, the phage or plasmids andaccompanying peptides were examined by ELISA to determine if thepeptides bind specifically to E-selectin. This assay was carried outsimilarly to the procedures used in the affinity enrichment process,except that after removing unbound phage (or plasmids), the wells weretypically treated with rabbit anti-phage antibody, then with alkalinephosphatase (AP)-conjugated goat anti-rabbit antibody, or withanti-LacI-AP, as shown in FIG. 3, part A, and then the amount ofalkaline phosphatase in each well was determined by standard methods.

By comparing test wells with control wells (no receptor), one candetermine whether a peptide binds to E-selectin specifically. Peptidesfound to bind specifically to E-selectin were then synthesized as thefree peptide (no phage) and tested in an HL60 cell adhesion competitionassay. The competition assay was carried out similarly to the ELISA,except that labeled HL60 cells were added in addition to the peptide[the control wells were typically of at least two types: (1) noimmobilized E-selectin; and (2) no peptide; see Example 1]. In thisregard, HL60 cells are known to contain surface glycoconjugates whichbind to E-selectin.

Peptides that block the binding of labeled HL60 cells to E-selectin arethen evaluated to determine their IC₅₀ -HL60 in the manner alsospecified in Example 1. Accordingly, the above methods provide in vitroprocedures for readily ascertaining peptides which bind to E-selectin aswell as determining the IC₅₀ -HL60 of such binding peptides in the assaydescribed in Example 1 and peptides which have an IC₅₀ -HL60 of 100 μMor less are deemed to bind to E-selectin.

Using this assay, the peptide HITWDQLWNVMN (SEQ ID NO 23) was determinedto have an IC₅₀ -HL60 of 50 μM and the peptide DITWDQLWDLMK (SEQ ID NO3) was determined to have an IC₅₀ -HL60 of 10 μM.

This latter peptide has strong binding affinity to E-selectin and,accordingly, was used as a standard in a competitive binding assay withcandidate peptides and peptide mimetics in order to determine relativebinding affinities for E-selectin. This competitive binding assay isconducted in the manner described in Example 2 hereinbelow and thatamount of peptide or peptide mimetic which inhibits by 50% the bindingof the peptide DITWDQLWDLMK (SEQ ID NO 3) to E-selectin is defined asthe IC₅₀ -standard for that compound. Peptides and peptide mimeticshaving an IC₅₀ -standard of about 2.5 mM or less define a preferred setof peptides and peptide mimetics having a greater binding affinity toE-selectin as measured by an IC₅₀ -HL60 of 100 μM.

The immobilized E-selectin used in the affinity enrichment and ELISAprocedures was produced in recombinant host cells in a truncated formcomprising the complete extracellular domain (amino acids 1 through554). The DNA encoding E-selectin is commercially available (R&DSystems, 614 McKinley Place NE, Minneapolis, Minn., 55413 USA as productcode BBG57). This truncated E-selectin molecule can be produced in avariety of different forms and host cells. One useful form of E-selectinis constructed by expressing the protein as a soluble protein inbaculovirus transformed host cells using standard methods; anotheruseful form is constructed with a signal peptide for protein secretionand for glycophospholipid membrane anchor attachment. This form ofanchor attachment is called "PIG-tailing;" see Caras and Weddell³³, andLin, et al.³⁴

Using the PIG-tailing system, one can cleave the E-selectin receptorfrom the surface of the cells expressing E-selectin (e.g., transformedCHO cells selected for high level expression of E-selectin with a cellsorter) with phospholipase C. The cleaved E-selectin still comprises acarboxy terminal sequence of amino acids, called the "HPAP tail," fromthe signal for membrane attachment and can be immobilized withoutfurther purification.

The recombinant receptor protein can be immobilized by coating the wellsof microtiter plates with an anti-HPAP tail antibody, blockingnon-specific binding with bovine serum albumin (BSA) in PBS, and thenbinding cleaved recombinant, truncated E-selectin to the antibody. Usingthis procedure, one should perform the immobilization reaction invarying concentrations of E-selectin to determine the optimum amount fora given preparation, because different preparations of recombinantprotein often contain different amounts of the desired protein. Inaddition, one should ensure that the immobilizing antibody is completelyblocked (with E-selectin or some other blocking compound) during theaffinity enrichment process. Otherwise, unblocked antibody can bindundesired phage during the affinity enrichment procedure. One can usepeptides that bind to the immobilizing antibody to block unbound sitesthat remain after ELAM-1 immobilization to avoid this problem or one cansimply immobilize the receptor directly to the wells of microtiterplates, without the aid of an immobilizing antibody. See U.S. patentapplication Ser. No. 07/947,339, filed Sep. 18, 1992, incorporatedherein by reference.

When using random peptide generation systems that allow for multivalentligand-receptor interaction, one must recognize that the density of theimmobilized receptor is an important factor in determining the affinityof the ligands that can bind to the immobilized receptor. At higherreceptor densities (e.g., each anti-receptor antibody-coated welltreated with 0.25 to 0.5 μg of receptor), multivalent binding is morelikely to occur than at lower receptor densities (e.g., eachanti-receptor antibody-coated well treated with 0.5 to 1 ng of thereceptor). If multivalent binding is occurring, then one will be morelikely to isolate ligands with relatively lower affinity, unless oneuses high densities of immobilized receptor to identify lead compoundsand uses lower receptor densities to isolate derivative compounds withhigher affinity for the receptor than the lead compounds.

Preferred screening methods to facilitate identification of peptideswhich bind E-selectin involve first identifying lead peptides which bindE-selectin and then making other peptides which resemble the leadpeptide. Specifically, using the pIII-based peptides on phage system, a12-mer library was screened to discover a phage that presents a peptidethat binds to E-selectin. The phage DNAs were sequenced to determine thesequences of the peptides displayed on the surface of the phages.Sequencing revealed that all phage displayed the same peptide. Thepeptide had the amino acid sequence: HITWDQLWNVMN (SEQ ID NO 23). Thispeptide can block the adhesion of HL60 cells to immobilized E-selectinand, as noted elsewhere, had an IC₅₀ -HL60 of 100 μM (see Example 1).

This peptide sequence serves as the basis for the construction ofanother peptide library designed to contain a high frequency ofderivatives of the peptide. Such a library can be synthesized so as tofavor the production of peptides that differ from the binding peptide inonly a few residues. This approach involves the synthesis (e.g.,chemical synthesis) of an oligonucleotide with the binding peptidecoding sequence, except that rather than using pure preparations of eachof the four nucleoside triphosphates in the synthesis, one uses mixturesof the four nucleoside triphosphates (i.e., 55% of the "correct"nucleotide, and 15% each of the other three nucleotides is one preferredmixture for this purpose and 70% of the "correct" nucleotide, and 10%each of the other three nucleotides is another preferred mixture forthis purpose) so as to generate derivatives of the binding peptidecoding sequence.

A variety of mutagenesis strategies were used to derivatize the leadpeptide by making "mutagenesis on a theme" libraries, which includedmutagenesis of the original coding sequence at 70:10:10:10 and55:15:15:15 frequencies; fixed-sliding libraries, such as HITWDQXXXXXX(SEQ ID NO 24), XXTWDQLWXXXX (SEQ ID NO 25), XXXXDQLWNVXX (SEQ ID NO26), and XXXXXXLWNVMN (SEQ ID NO 27); an extended/mutagenized library,XXXXHITWDQLWNVMNXXXX (70:10:10:10) (SEQ ID NO 28), which was screenedusing standard and modified [200 μM of E-selectin binding peptideQITWAQLWNMMK (SEQ ID NO 7) for 2 hours, followed by glycine-HCl] elutionconditions; and libraries with fixed amino acids, such as XXXWXXLWXXMX(SEQ ID NO 29) and XXXXWXXLWXXMXXXXXX (SEQ ID NO 30) produced in apeptides-on-plasmids system and tested under standard and low receptordensity conditions. Screening such libraries yielded theE-selectin-binding peptides shown in Table 1 below.

                                      TABLE 1                                     __________________________________________________________________________    HITWDQLWNVMN (SEQ ID NO 23)                                                                       AITWDQLWLLMS (SEQ ID NO 31)                               ELTWDQLWVLMS (SEQ ID NO 32)                                                                       DVTWDQLWELMT (SEQ ID NO 33)                               EVTWDQLWVMMQ (SEQ ID NO 34)                                                                       NLTWDQLWVLMS (SEQ ID NO 35)                               EMSWLELWNVMN (SEQ ID NO 36)                                                                       TITWDQLWQMMS (SEQ ID NO 37)                               ELSWDQLWNVMN (SEQ ID NO 38)                                                                       EMTWQELWNVMN (SEQ ID NO 39)                               EMTWTELWNVMN (SEQ ID NO 40)                                                                       DMTWSQLWNVMN (SEQ ID NO 41)                               EMTWLGLWNVMN (SEQ ID NO 42)                                                                       QITWMELWNLMN (SEQ ID NO 43)                               EITWDQLWEVMN (SEQ ID NO 44)                                                                       EITWDQLWDVMN (SEQ ID NO 45)                               DISWDQLWNVMN (SEQ ID NO 46)                                                                       QITWDQLWDLMK (SEQ ID NO 47)                               EMTWDQLWNVMN (SEQ ID NO 48)                                                                       DITWDQLWNMMD (SEQ ID NO 49)                               DITWNMLWNMMQ (SEQ ID NO 50)                                                                       DISWDDLWIMMN (SEQ ID NO 51)                               DITWHQLWNLMN (SEQ ID NO 52)                                                                       EISWEQLWTMMN (SEQ ID NO 53)                               DITWEQLWNMMN (SEQ ID NO 54)                                                                       EITWDQLWTLMT (SEQ ID NO 55)                               DITWHQLWNLMN (SEQ ID NO 56)                                                                       DMTWDQLWIVMN (SEQ ID NO 57)                               DITWEQLWNLMN (SEQ ID NO 58)                                                                       QITWYQLWNMMN (SEQ ID NO 59)                               HISWHELWNLMQ (SEQ ID NO 60)                                                                       YITWEQLWTMMN (SEQ ID NO 61)                               HITWDQLWDLMQ (SEQ ID NO 62)                                                                       QITWDQLWDLMY (SEQ ID NO 63)                               QITWDQLWNMMI (SEQ IO NO 64)                                                                       QITWAQLWNMMK (SEQ ID NO 7)                                YITWEQLWNMMN (SEQ ID NO 65)                                                                       HITWDQLWDIMS (SEQ ID NO 66)                               HITWDQLWEIMS (SEQ ID NO 67)                                                                       HITWDQLWALMT (SEQ ID NO 68)                               HITWDQLWSLMS (SEQ ID NO 69)                                                                       HITWDQLWLMMS (SEQ ID NO 70)                               HITWDQLWDLMQ (SEQ ID NO 71)                                                                       HITWDQLWWIMA (SEQ ID NO 72)                               HITWDQLWLLMA (SEQ ID NO 73)                                                                       HITWDQLWMLMA (SEQ ID NO 74)                                      GSDSHITWDELWNLMNPVLA (SEQ ID NO 75)                                           NWLDDITWDELWKIMNPSTA (SEQ ID NO 76)                                           ETDDHITWDQLWRIMTATMA (SEQ ID NO 77)                                           WTDTHITWDQLWHFMNMGEQ (SEQ ID NO 78)                                           GFGEAITWDQLWDMMNGEDA (SEQ ID NO 79)                                           NVAEQITWDQLWNLMSVGSS (SEQ ID NO 80)                                           GQTGLITWDMLWNLMNPVGE (SEQ ID NO 81)                                           GTGDHITWDQLWNLMINEKG (SEQ ID NO 82)                                           EYGRHITWDQLWQLMQSATA (SEQ ID NO 83)                                           MNNWHVSWEQLWDIMNGPPN (SEQ ID NO 84)                                           ESASHITWGQLWDLMNASEV (SEQ ID NO 85)                                           YWRGNITWDQLWNIMNSEYS (SEQ ID NO 86)                                           AGASHITWAQLWNMMNGNEG (SEQ ID NO 87)                                           GSWAHITWDQLWNLMNMGTQ (SEQ ID NO 88)                                           YGNSNITWDQLWSIMNRQTT (SEQ ID NO 89)                                           AHLPHISWDTLWHIMNKGEK (SEQ ID NO 90)                                           ESASHITWGQLWDLMNASEV (SEQ ID NO 91)                                           MNNWHVSWEQLWDIMNGPPN (SEQ ID NO 92)                                           GFGEAITWDQLWDMMNGEDA (SEQ ID NO 93)                                           WTDTHITWDQLWHFMNMGEQ (SEQ ID NO 94)                                    EMTWAELWTLME (SEQ ID NO 95)                                                                       DYTWFELWDMMQ (SEQ ID NO 5)                                DYSWHDLWEMMS (SEQ ID NO 8)                                                                        DISWRQLWDIMN (SEQ ID NO 96)                               EISWLGLWDIMN (SEQ ID NO 97)                                                                       DMTWHDLWTLMS (SEQ ID NO 98)                               RGVWGGLWSMTW (SEQ ID NO 99)                                                                       EMTWQQLWVVMQ (SEQ ID NO 100)                                     AEWTWDQLWHVMNPAESQ (SEQ ID NO 101)                                            RNMSWLELWEHMK (SEQ ID NO 102)                                                 SQVTWNDLWSVMNPEVVN (SEQ ID NO 103)                                            HRAEWLALWEQMSP (SEQ ID NO 104)                                                YKKEWLELWHQMQA (SEQ ID NO 105)                                                RSLSWLQLWDQMK (SEQ ID NO 106)                                                 KEQQWRNLWKMMS (SEQ ID NO 107)                                                 KKEDWLALWRIMSVPD (SEQ ID NO 108)                                              RNMSWLELWEHMK (SEQ ID NO 109)                                                 GRPTWNELWDMMQAP (SEQ ID NO 110)                                               KRKQWIELWNIMS (SEQ ID NO 111)                                                 KTSEWNNLWKLMSQ (SEQ ID NO 112)                                         __________________________________________________________________________

The norleucine derivatives, in which the conserved M is replaced withNle, are especially noteworthy, as these derivatives may be more stableunder certain environmental conditions than other peptides of theinvention that contain an M at this position. Examples of such preferredpeptides wherein M is replaced by Nle are as follows:

DITWDQLWDLNleK (SEQ ID NO 9)

DGDITWDQLWDLNleK (SEQ ID NO 113)

QITWDQLWDLNleK (SEQ ID NO 115)

AITWDQLWDLNleK-OH (SEQ ID NO 134)

DATWDQLWDLNleK-OH (SEQ ID NO 135)

DITADQLWDLNleK-OH (SEQ ID NO 136)

DITWAQLWDLNleK-OH (SEQ ID NO 137)

DITWDALWDLNleK-OH (SEQ ID NO 138)

DITWDQAWDLNleK-OH (SEQ ID NO 139)

DITWDQLADLNleK-OH (SEQ ID NO 140)

DITWDQLWALNleK-OH (SEQ ID NO 141)

DITWDQLWDANleK-OH (SEQ ID NO 142)

DITWDQLWDLAK-OH (SEQ ID NO 143)

DITWDQLWDLNleA-OH (SEQ ID NO 144)

Ac-DITWDQLWDLNleK-NH₂ (SEQ ID NO 158)

Peptides and peptide mimetics having an IC₅₀ -HL60 of greater than about100 μM lack sufficient binding to permit use in either the diagnostic ortherapeutic aspects of this invention. Preferably, for diagnosticpurposes, the peptides and peptide mimetics have an IC₅₀ -standard ofabout 2.5 mM or less and, for pharmaceutical purposes, the peptides andpeptide mimetics have and IC₅₀ -standard of about 2 mM or less.

The binding peptide sequence also provides a means to determine theminimum size of an E-selectin binding, cell adhesion blocking compoundof the invention. Several deletion analogs of a lead peptide of theinvention are shown in FIG. 4; one preferred peptide is ITWDQLWDLMK SEQID NO 160). Using the "very large scale immobilized polymer synthesis"system described in U.S. Pat. No. 5,143,845 and related publications,supra, one not only can determine the minimum size of a peptide withE-selectin binding activity, but also can make all of the peptides thatform the group of peptides that differ from the lead compound in one,two, or more residues, or that differ in containing non-natural aminoacids. This collection of peptides and peptide mimetics can then bescreened for ability to bind to E-selectin. This immobilized polymersynthesis system or other peptide synthesis methods, such as the ESLmethod discussed above, can also be used to synthesize every truncation,deletion, and every combination of truncation and deletion analogs ofthe peptide compounds of the invention.

Random peptide synthesis can also be employed to generate a multiplicityof other peptides of varying length which can be used in the assaymethods described herein to determine E-selectin binding. Once peptideswhich bind E-selectin are determined, these peptides can be used as"lead peptides" in the manner described above to obtain peptides havingenhanced E-selectin binding.

Additionally, the following peptides were identified by the randompeptide diversity generation systems described above with the exceptionthat the expression was on pVIII instead of pIII for identificationpurposes. Several of these peptides were actually found in a pIIImutagenesis library but others (SEQ ID NOS 11-15) were found only onpVIII.

YDDVCCELLF (SEQ ID NO 11)

DLPQWYTEWC (SEQ ID NO 12)

ENSHWCTCPC (SEQ ID NO 13)

DIEQDWVTWM (SEQ ID NO 14)

NEWCVVPCRL (SEQ ID NO 15)

DIWQDWVRWM (SEQ ID NO 16)

DLWQDWVTWM (SEQ ID NO 17)

DLWQDWVHWM (SEQ ID NO 18)

DIWQDWVTWM (SEQ ID NO 19)

DIWQDWVKWM (SEQ ID NO 20)

DIWQDWVRWM-NH₂ (SEQ ID 16)

DIWEDWVRWM (SEQ ID 21)

DIWQDWITWM (SEQ ID 22)

The simple in vitro assays set forth above and in the exampleshereinbelow provide a straightforward method for preparing and screeningpeptides and peptide mimetics for their binding properties toE-selectin.

B. Preparation of Peptides and Peptide Mimetics Which Bind E-SelectinPeptides

The peptides identified above as binding E-selectin can also be preparedby classical methods known in the art including standard solid phasetechniques. The standard methods include exclusive solid phasesynthesis, partial solid phase synthesis, fragment condensation,classical solution synthesis, and recombinant DNA technology. See, e.g.,Merrifield³⁵. On solid phase, the synthesis is typically commenced fromthe C-terminal end of the peptide using an α-amino protected resin. Asuitable starting material can be prepared, for instance, by attachingthe required α-amino acid to a chloromethylated resin, a hydroxymethylresin, or a benzhydrylamine resin. One such chloromethylated resin issold under the tradename BIO-BEADS SX-1 by BioRad Laboratories,Richmond, Calif., USA, and the preparation of the hydroxymethyl resin isdescribed by Bodonszky, et al.³⁶ The benzhydrylamine (BHA) resin hasbeen described by Pietta and Marshall³⁷, and is commercially availablefrom Beckman Instruments, Inc., Palo Alto, Calif., USA, in thehydrochloride form. A particularly preferred resin is a PAL resinavailable from Millipore Corp., Bedford, Mass., USA.

Thus, the compounds of the invention can be prepared by coupling anα-amino protected amino acid to the chloromethylated resin with the aidof, for example, cesium bicarbonate catalyst, according to the methoddescribed by Gisin³⁸. After the initial coupling, the α-amino protectinggroup is removed by a choice of reagents including trifluoroacetic acid(TFA) or hydrochloric acid (HCl) solutions in organic solvents at roomtemperature.

The α-amino protecting groups are those known to be useful in the art ofstepwise synthesis of peptides. Included are acyl type protecting groups(e.g. formyl, trifluoroacetyl, acetyl), aromatic urethane typeprotecting groups (e.g. benzyloxycarboyl (Cbz) and substituted Cbz),aliphatic urethane protecting groups (e.g. t-butyloxycarbonyl (Boc),isopropyloxycarbonyl, cyclohexyloxycarbonyl, 9-fluorenyl-methoxycarbonyl(F-moc) and alkyl type protecting groups (e.g. benzyl, triphenylmethyl).Boc and Fmoc are preferred protecting groups. The side chain protectinggroup remains intact during coupling and is not split off during thedeprotection of the amino-terminus protecting group or during coupling.The side chain protecting group must be removable upon the completion ofthe synthesis of the final peptide and under reaction conditions thatwill not alter the target peptide.

The side chain protecting groups for Tyr include tetrahydropyranyl,tert-butyl, trityl, benzyl, Cbz, 2-Br-Cbz, and 2,5-dichlorobenzyl. Theside chain protecting groups for Asp include benzyl, 2,6-dichlorobenzyl,methyl, ethyl, and cyclohexyl. The side chain protecting groups for Thrand Ser include acetyl, benzoyl, trityl, tetrahydropyranyl, benzyl,2,6-dichlorobenzyl, and Cbz. The side chain protecting groups for Thrand Ser is benzyl. The side chain protecting groups for Arg includenitro, tosyl (Tos), Cbz, adamantyloxycarbonyl mesitoylsulfonyl (Mts), orBoc. The side chain protecting groups for Lys include Cbz,2-chlorobenzyloxy-carbonyl (2-Cl -Cbz), 2-bromobenzyloxycarbonyl(2-Br-Cbz), Tos, or Boc.

After removal of the α-amino protecting group, the remaining protectedamino acids are coupled stepwise in the desired order. Each protectedamino acid is generally reacted in about a 3-fold excess using anappropriate carboxyl group activator such as dicyclohexylcarbodiimide(DCC) in solution, for example, in methylene chloride (CH₂ Cl₂),dimethyl formamide (DMF) mixtures.

After the desired amino acid sequence has been completed, the desiredpeptide can be decoupled from the resin support by treatment with areagent such as hydrogen fluoride (HF) or trifluoroacetic acid (TFA),which not only cleaves the peptide from the resin, but also cleaves allremaining side chain protecting groups. When the chloromethylated resinis used, hydrogen fluoride treatment results in the formation of thefree peptide acids. When the benzhydrylamine resin is used, hydrogenfluoride treatment results directly in the free peptide amide (i.e., theC-terminus is --C(O)NH₂). Alternatively, when the chloromethylated resinis employed, the side chain protected peptide can be decoupled bytreatment of the peptide resin with ammonia to give the desired sidechain protected amide or with an alkylamine to give a side chainprotected alkylamide or dialkylamide (i.e., the C-terminus is --C(O)NR³R⁴ where R³ and R⁴ are as defined above). Side chain protection is thenremoved in the usual fashion by treatment with hydrogen fluoride to givethe free amides, alkylamides, or dialkylamides.

These solid phase peptide synthesis procedures are well known in the artand further described in Stewart⁴⁰.

The amino acids employed in the above described methods for peptidesynthesis can either be naturally occurring amino acids or syntheticamino acids other than the 20 naturally occurring, genetically encodedamino acids. For instance, naphthylalanine can be substituted fortryptophan, facilitating synthesis. Other synthetic amino acids that canbe substituted into the peptides of the present invention includeL-hydroxypropylglycine, L-3,4-dihydroxyphenylalanyl, α-amino acids suchas L-α-hydroxylysyl and D-α-methylalanyl, L-α-methyl-alanyl, βamino-acids such as β-analine, and isoquinolyl.

D-amino acids and other non-naturally occurring synthetic amino acidscan also be incorporated into the peptides of the present invention.Such other non-naturally occurring synthetic amino acids include thosewhere the naturally occurring side chains of the 20 genetically encodedamino acids (or any L or D amino acid) are replaced with other sidechains, for instance with groups such as alkyl, lower alkyl, cyclic 4-,5-, 6-, to 7-membered alkyl, amide, amide lower alkyl, amide di(loweralkyl), lower alkoxy, hydroxy, carboxy and the lower ester derivativesthereof, and with 4-, 5-, 6-, to 7-membered hetereocyclic. Inparticular, proline analogs in which the ring size of the prolineresidue is changed from 5 members to 4, 6, or 7 members can be employed.

Cyclic groups can be saturated or unsaturated, and if unsaturated, canbe aromatic or non-aromatic. Heterocyclic groups preferably contain oneor more nitrogen, oxygen, and/or sulphur heteroatoms. Examples of suchgroups include the furazanyl, furyl, imidazolidinyl, imidazolyl,imidazolinyl, isothiazolyl, isoxazolyl, morpholinyl (e.g. morpholino),oxazolyl, piperazinyl (e.g. 1-piperazinyl), piperidyl (e.g. 1-piperidyl,piperidino), pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl,pyridazinyl, pyridyl, pyrimidinyl, pyrrolidinyl (e.g. 1-pyrrolidinyl),pyrrolinyl, pyrrolyl, thiadiazolyl, thiazolyl, thienyl, thiomorpholinyl(e.g. thiomorpholino), and triazolyl. These heterocyclic groups can besubstituted or unsubstituted. Where a group is substituted, thesubstituent can be alkyl, alkoxy, halogen, oxygen, or substituted orunsubstituted phenyl.

One can also readily modify peptides which bind E-selectin byphosphorylation, and other methods for making peptide derivatives of thecompounds of the present invention are described in Hruby et al.⁴² Thus,the peptide compounds of the invention also serve as a basis to preparepeptide mimetics with similar biological activity.

Peptide Mimetics

Those of skill in the art recognize that a variety of techniques areavailable for constructing peptide mimetics with the same or similardesired biological activity as the corresponding peptide compound butwith more favorable activity than the peptide with respect tosolubility, stability, and susceptibility to hydrolysis and proteolysis.See, for example, Morgan and Gainor⁴¹. The following describes methodsfor preparing peptide mimetics modified at the N-terminal amino group,the C-terminal carboxyl group, and/or changing one or more of the amidolinkages in the peptide to a non-amido linkage. It being understood thattwo or more such modifications can be coupled in one peptide mimeticstructure (e.g., modification at the C-terminal carboxyl group andinclusion of a --CH₂ -carbamate linkage between two amino acids in thepeptide).

Modification of the N-amino Terminus

After the desired amino acid sequence has been completed as describedabove, the blocking group on the N-terminus amino group can beselectively removed so as to provide for a peptide sequence blocked atall positions other than the N-terminal amino group and attached to asolid resin through the C-terminus. One can then modify the aminoterminus of the peptides of the invention to produce peptide mimetics ofthe invention.

Amino terminus modifications are as recited above and includealkylating, acetylating, adding a carbobenzoyl group, forming asuccinimide group, etc. Specifically, the N-terminal amino group canthen be reacted as follows:

(a) to form an amide group of the formula RC(O)NH-- where R is asdefined above by reaction with an acid halide [e.g., RC(O)Cl] or acidanhydride. Typically, the reaction can be conducted by contacting aboutequimolar or excess amounts (e.g., about 5 equivalents) of an acidhalide to the peptide in an inert diluent (e.g., dichloromethane)preferably containing an excess (e.g., about 10 equivalents) of atertiary amine, such as diisopropylethylamine, to scavenge the acidgenerated during reaction. Reaction conditions are otherwiseconventional (e.g., room temperature for 30 minutes). Alkylation of theterminal amino to provide for a lower alkyl N-substitution followed byreaction with an acid halide as described above will provide for N-alkylamide group of the formula RC(O)NR--;

(b) to form a succinimide group by reaction with succinic anhydride. Asbefore, an approximately equimolar amount or an excess of succinicanhydride (e.g., about 5 equivalents) can be employed and the aminogroup is converted to the succinimide by methods well known in the artincluding the use of an excess (e.g., ten equivalents) of a tertiaryamine such as diisopropylethylamine in a suitable inert solvent (e.g.,dichloromethane). See, for example, Wollenberg, et al., U.S. Pat. No.4,612,132 which is incorporated herein by reference in its entirety. Itis understood that the succinic group can be substituted with, forexample, C₂ -C₆ alkyl or --SR substituents which are prepared in aconventional manner to provide for substituted succinimide at theN-terminus of the peptide. Such alkyl substituents are prepared byreaction of a lower olefin (C₂ -C₆) with maleic anhydride in the mannerdescribed by Wollenberg, et al., supra. and --SR substituents areprepared by reaction of RSH with maleic anhydride where R is as definedabove;

(c) to form a benzyloxycarbonyl-NH--or a substitutedbenzyloxycarbonyl-NH-- group by reaction with approximately anequivalent amount or an excess of CBZ-Cl (i.e., benzyloxycarbonylchloride) or a substituted CBZ-Cl in a suitable inert diluent (e.g.,dichloromethane) preferably containing a tertiary amine to scavenge theacid generated during the reaction;

(d) to form a sulfonamide group by reaction with an equivalent amount oran excess (e.g., 5 equivalents) of R--S(O)₂ Cl in a suitable inertdiluent (dichloromethane) to convert the terminal amine into asulfonamide where R is as defined above. Preferably, the inert diluentcontains excess tertiary amine (e.g., ten equivalents) such asdiisopropylethylamine, to scavenge the acid generated during reaction.Reaction conditions are otherwise conventional (e.g., room temperaturefor 30 minutes);

(e) to form a carbamate group by reaction with an equivalent amount oran excess (e.g., 5 equivalents) of R--OC(O)Cl or R--OC(O)OC₆ H₄ --p--NO₂in a suitable inert diluent (e.g., dichloromethane) to convert theterminal amine into a carbamate where R is as defined above. Preferably,the inert diluent contains an excess (e.g., about 10 equivalents) of atertiary amine, such as diisopropylethylamine, to scavenge any acidgenerated during reaction. Reaction conditions are otherwiseconventional (e.g., room temperature for 30 minutes); and

(f) to form a urea group by reaction with an equivalent amount or anexcess (e.g., 5 equivalents) of R--N═C═O in a suitable inert diluent(e.g., dichloromethane) to convert the terminal amine into a urea (i.e.,RNHC(O)NH--) group where R is as defined above. Preferably, the inertdiluent contains an excess (e.g., about 10 equivalents) of a tertiaryamine, such as diisopropylethylamine. Reaction conditions are otherwiseconventional (e.g., room temperature for about 30 minutes).

As shown in the examples below, peptides having modifications to theN-terminus amino group provide for peptide mimetics having bindingproperties to E-selectin as determined by an IC₅₀ -standard of less than2.5 mM.

Modification of the C-Terminus

In preparing peptide mimetics wherein the C-terminal carboxyl group isreplaced by an ester (i.e., --C(O)OR where R is as defined above), theresins used to prepare the peptide acids are employed, and the sidechain protected peptide is cleaved with base and the appropriatealcohol, e.g., methanol. Side chain protecting groups are then removedin the usual fashion by treatment with hydrogen fluoride to obtain thedesired ester.

In preparing peptide mimetics wherein the C-terminal carboxyl group isreplaced by the amide --C(O)NR³ R⁴, a benzhydrylamine resin is used asthe solid support for peptide synthesis. Upon completion of thesynthesis, hydrogen fluoride treatment to release the peptide from thesupport results directly in the free peptide amide (i.e., the C-terminusis --C(O)NH₂). Alternatively, use of the chloromethylated resin duringpeptide synthesis coupled with reaction with ammonia to cleave the sidechain protected peptide from the support yields the free peptide amideand reaction with an alkylamine or a dialkylamine yields a side chainprotected alkylamide or dialkylamide (i.e., the C-terminus is --C(O)NRR¹where R and R¹ are as defined above). Side chain protection is thenremoved in the usual fashion by treatment with hydrogen fluoride to givethe free amides, alkylamides, or dialkylamides.

In another alternative embodiment, the C-terminal carboxyl group or aC-terminal ester can be induced to cyclize by internal displacement ofthe --OH or the ester (--OR) of the carboxyl group or ester respectivelywith the N-terminal amino group to form a cyclic peptide. For example,after synthesis and cleavage to give the peptide acid, the free acid isconverted to an activated ester by an appropriate carboxyl groupactivator such as dicyclohexylcarbodiimide (DCC) in solution, forexample, in methylene chloride (CH₂ Cl₂), dimethyl formamide (DMF)mixtures. The cyclic peptide is then formed by internal displacement ofthe activated ester with the N-terminal amine. Internal cyclization asopposed to polymerization can be enhanced by use of very dilutesolutions. Such methods are well known in the art.

As shown in the examples below, peptides having modifications to theC-terminal carboxyl group provide for peptide mimetics having bindingproperties to E-selectin as determined by an IC₅₀ -standard of less than2.5 mM.

Modification to Incorporate a Non-Peptidyl Linkage

Peptide mimetics wherein one or more of the peptidyl linkages[--C(O)NH--] have been replaced by such linkages as a --CH₂ -carbamatelinkage, a phosphonate linkage, a --CH₂ -sulfonamide linkage, a urealinkage, a secondary amine (--C₂ NH--) linkage, and an alkylatedpeptidyl linkage [--C(O)NR⁶ -- where R⁶ is lower alkyl] are preparedduring conventional peptide synthesis by merely substituting a suitablyprotected amino acid analogue for the amino acid reagent at theappropriate point during synthesis.

Suitable reagents include, for example, amino acid analogues wherein thecarboxyl group of the amino acid has been replaced with a moietysuitable for forming one of the above linkages. For example, if onedesires to replace a --C(O)NR-- linkage in the peptide with a --CH₂-carbamate linkage (--CH₂ OC(O)NR--), then the carboxyl (--COOH) groupof a suitably protected amino acid is first reduced to the --CH₂ OHgroup which is then converted by conventional methods to a --OC(O)Clfunctionality or a para-nitrocarbonate --OC(O)O--C₆ H₄ --p--NO₂functionality. Reaction of either of such functional groups with thefree amine or an alkylated amine on the N-terminus of the partiallyfabricated peptide found on the solid support leads to the formation ofa --CH₂ OC(O)NR-- linkage. For a more detailed description of theformation of such --CH₂ -carbamate linkages, see Cho, et al.³⁹

Similarly, replacement of an amido linkage in the peptide with aphosphonate linkage can be achieved in the manner set forth in U.S.patent application Ser. Nos. 07/943,805, 08/081,577, and 08/119,700, thedisclosures of which are incorporated herein by reference in theirentirety.

Replacement of an amido linkage in the peptide with a --CH₂ -sulfonamidelinkage can be achieved by reducing the carboxyl (--COOH) group of asuitably protected amino acid to the --CH₂ OH group and the hydroxylgroup is then converted to a suitable leaving group such as a tosylgroup by conventional methods. Reaction of the tosylated derivativewith, for example, thioacetic acid followed by hydrolysis and oxidativechlorination will provide for the --CH₂ --S(O)₂ Cl functional groupwhich replaces the carboxyl group of the otherwise suitably protectedamino acid. Use of this suitably protected amino acid analogue inpeptide synthesis provides for inclusion of an --CH₂ S(O)₂ NR-- linkagewhich replaces the amido linkage in the peptide thereby providing apeptide mimetic. For a more complete description on the conversion ofthe carboxyl group of the amino acid to a --CH₂ S(O)₂ Cl group, see, forexample, Chemistry & Biochemistry of Amino Acids, Peptides and Proteins,Edited by Boris Weinstein, Vol. 7, pp. 267-357, Marcel Dekker, Inc., NewYork, N.Y. (1983) which is incorporated herein by reference in itsentirety.

Replacement of an amido linkage in the peptide with a urea linkage canbe achieved in the manner set forth in U.S. patent application Ser. No.08/147,805 which application is incorporated herein by reference in itsentirety.

Secondary amine linkages wherein a --CH₂ NH-- linkage replaces the amidolinkage in the peptide can be prepared by employing, for example, asuitably protected dipeptide analogue wherein the carbonyl bond of theamido linkage has been reduced to a CH₂ group by conventional methods.For example, in the case of diglycine, reduction of the amide to theamine will yield after deprotection H₂ NCH₂ CH₂ NHCH₂ COOH which is thenused in N-protected form in the next coupling reaction. The preparationof such analogues by reduction of the carbonyl group of the amidolinkage in the dipeptide is well known in the art.

The suitably protected amino acid analogue is employed in theconventional peptide synthesis in the same manner as would thecorresponding amino acid. For example, typically about 3 equivalents ofthe protected amino acid analogue are employed in this reaction. Aninert organic diluent such as methylene chloride or DMF is employed and,when an acid is generated as a reaction by-product, the reaction solventwill typically contain an excess amount of a tertiary amine to scavengethe acid generated during the reaction. One particularly preferredtertiary amine is diisopropylethylamine which is typically employed inabout 10 fold excess. The reaction results in incorporation into thepeptide mimetic of an amino acid analogue having a non-peptidyl linkage.Such substitution can be repeated as desired such that from zero to allof the amido bonds in the peptide have been replaced by non-amido bonds.

Utility

The peptides and peptide mimetics of the invention are useful in vitroas unique tools for understanding the biological role of E-selectin, aswell as other receptors of the selectin family, such as CD62, that maybind to the peptide compounds of the invention. The peptides and peptidemimetics of the invention can also be used to evaluate the many factorsthought to influence, and be influenced by, the cell adhesion process.The present compounds are also useful in the development of othercompounds that bind to E-selectin, because the present compounds provideimportant SAR information that facilitate that development.

The peptides and peptide mimetics of this invention can also be labeledwith a detectable label and used to diagnose the in vivo site(s) ofinflammation. When used for this purpose, a suitable detectable label iscovalently attached to a peptide or peptide mimetic of this inventionand an effective amount of the so-labeled peptide or peptide mimetic isadministered to the patient. Sufficient time after administration isallowed to pass so as to permit circulation of the labeled peptide orpeptide mimetic in the patient and attachment to the site of E-selectinexpression. At this point, detection of the label and its location inthe patient is determined so as to provide for the site of inflammation.

The peptides and peptide mimetics of this invention can also be employedto target conventional anti-inflammatory drugs or other agents tospecific sites of tissue injury. By using an E-selectin binding peptideor peptide mimetic of this invention to target such a drug to theE-selectin receptor on, e.g., a vascular endothelial cell, suchdrug/peptide composites can achieve higher drug concentrations at thesites of injury. Side effects from the conventional administration ofanti-inflammatory drugs can be reduced by the lower dosages of drugrequired and the localization of the drug at the site of injury.

Preferably, the anti-inflammatory drug is coupled to the peptide orpeptide mimetic of this invention via covalent linkages and is conductedin a manner such the coupling does not substantially inhibit the abilityof the peptide or peptide mimetic from binding to E-selectin. A varietyof anti-inflammatory agents can be coupled for targetting in this mannerincluding, by way of example, cyclosporin A, indomethacin, naproxen,FK-506, mycophenolic acid, etc.

The peptides and peptide mimetics of the invention can also beadministered to warm blooded animals, including humans, to treatconditions mediated by E-selectin expression in vivo. Thus, the presentinvention encompasses methods for therapeutic treatment of E-selectinrelated disorders, especially inflammatory disorders, that compriseadministering a peptide or peptide mimetic of the invention in amountssufficient to block the adhesion of leukocytes to E-selectin in vivo.For example, the peptides and compounds of the invention can beadministered to control inflammatory conditions and other conditionsmediated by E-selectin. Such inflammatory conditions mediated byselectin proteins include, by way of example, (1) dermal inflammation,including Il-1-induced skin inflammation; atopic dermatitis, psoriasis,and allergic contact dermatitis; delayed-type hypersensitivity reaction;and endotoxin-induced acute cutaneous inflammation in baboons; (2)intestinal inflammation, including inflamed colonic mucosa of patientswith ulcerative or Crohn's colitis; inflammatory bowel disease; andintestinal graft-versus-host disease; (3) other sites of inflammation,including synovial tissues from patients with rheumatoid arthritis andosteo arthritis; rat endotoxin-induced uveitis, and inflamed endotheliumof asthma model (primate antigen inhalation), acute respiratory distresssyndome (ARDS); (4) septic shock, and (5) and the overproduction ofcytokines that stimulate expression of E-selectin, or other members ofthe selectin family.

In addition to the above human uses, it is contemplated that thesepeptides can be used in veterinary uses due to cross-reactivity with theselectins expressed in the particular animal.

Accordingly, the present invention includes pharmaceutical compositionscomprising, as an active ingredient, at least one of the peptides orpeptide mimetics of the invention in association with a pharmaceuticalcarrier or diluent. The peptides and peptide mimetics of this inventionare effective in treating E-selectin mediated conditions whenadministered at a dosage range of from about 0.001 mg to about 20 mg/kgof body weight per day. The specific dose employed is regulated by theparticular condition being treated, the route of administration as wellas by the judgement of the attending clinician depending upon factorssuch as the severity of the condition, the age and general condition ofthe patient, and the like.

The peptide and peptide mimetics of the invention can be administered byoral, parenteral (intramuscular, intraperitoneal, intravenous (IV) orsubcutaneous injection), transdermal, nasal, vaginal, rectal, pulmonary,sublingual or inhalation (via a fine powder formulation) routes ofadministration and can be formulated in dosage forms appropriate foreach route of administration.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activecompound is admixed with at least one inert pharmaceutically acceptablecarrier such as sucrose, lactose, or starch. Such dosage forms can alsocomprise, as is normal practice, additional substances other than inertdiluents, e.g., lubricating agents such as magnesium stearate. In thecase of capsules, tablets, and pills, the dosage forms may also comprisebuffering agents. Tablets and pills can additionally be prepared withenteric coatings.

Liquid dosage forms for oral administration include pharmaceuticallyacceptable emulsions, solutions, suspensions, syrups, with the elixirscontaining inert diluents commonly used in the art, such as water.Besides such inert diluents, compositions can also include adjuvants,such as wetting agents, emulsifying and suspending agents, andsweetening, flavoring, and perfuming agents.

Preparations according to this invention for parenteral administrationinclude sterile aqueous or non-aqueous solutions, suspensions, oremulsions. Examples of non-aqueous solvents or vehicles are propyleneglycol, polyethylene glycol, vegetable oils, such as olive oil and cornoil, gelatin, and injectable organic esters such as ethyl oleate. Suchdosage forms may also contain adjuvants such as preserving, wetting,emulsifying, and dispersing agents. They may be sterilized by, forexample, filtration through a bacteria retaining filter, byincorporating sterilizing agents into the compositions, by irradiatingthe compositions, or by heating the compositions. They can also bemanufactured using sterile water, or some other sterile injectablemedium, immediately before use.

Compositions for rectal or vaginal administration are preferablysuppositories which may contain, in addition to the active substance,excipients such as cocoa butter or a suppository wax. Compositions fornasal or sublingual administration are also prepared with standardexcipients well known in the art.

As can be appreciated from the disclosure above, the present inventionhas a wide variety of applications. Accordingly, the following examplesare offered by way of illustration, not by way of limitation.

In the examples below, the following abbreviations have the followingmeanings. If not defined below, then the abbreviations have their artrecognized meanings.

    ______________________________________                                        BSA       =       bovine serum albumin                                        DMEM      =       Dulbecco's modified eagle medium                            DMSO      =       dimethylsulfoxide                                           ELAM-1    =       endothelial leukocyte adhesion                                                molecule-1                                                  FCS       =       fetal calf serum                                            F-moc     =       9-fluorenyl-methoxycarbonyl                                 F12       =       F-12 nutrient mixture available from                                          Gibco                                                       HEPES     =       N-2-hydroxyethylpiperazine-N'-2-                                              ethanesulfonic acid                                         mL        =       milliliter                                                  mM        =       millimolar                                                  nM        =       nanomolar                                                   PBS       =       phosphate buffered saline                                   RPM       =       rotations per minute                                        SLe.sup.x =       sialyl Lewis.sup.x, i.e., αNeu5Ac(2→3)-                          Gal(1→4)-[α-L-Fuc(1→3)]-βGlcNAc                      -                                                           μCi    =       microCurie                                                  μg     =       micrograms                                                  μL     =       microliters                                                 μM     =       micromolar                                                  ______________________________________                                    

EXAMPLE 1 HL60 Cell Adhesion Assay

This example describes the results of a cell adhesion assay in which theability of HL60 cells (a promyelocytic leukemia-60 cell clone thatexpresses Sle^(x) and is available from the ATCC (Rockville, Md.) asATCC No. CCL 240) to adhere to immobilized E-selectin was tested in thepresence of various compounds, including an E-selectin binding peptideof the invention.

The assay requires HL60 cells labeled with tritiated thymidine. To labelthe cells, about 10 million HL60 cells are collected by centrifugation(2500 RPM for 3 minutes) and resuspended in 15 mL of DMEM/F12 plus 5%fetal calf serum containing 2 μCi/mL of tritiated thymidine (Amersham).The cells are then placed in a T75 flask and incubated at 37° C. for 24hours. The cells are then collected by centrifugation, washed twice withRPMI/HEPES/0.1% FCS, and resuspended in the same solution at aconcentration of about one to two million cells per mL. About 100 μL ofthis cell preparation are used per well of a 96-well microtiter dish inthe assay.

The wells of the microtiter dish were first coated with theanti-receptor immobilizing antibody, an anti-HPAP-tail antibody, usingabout 2.5 μg of antibody per well. The wells were then treated with asolution composed of 1% BSA in PBS and then with a 1:10 dilution (1:100and 1:500 dilutions and immobilizations were also conducted and tested)of a soluble E-selectin harvest (the E-selectin harvest, prepared bytreating cells expressing PIG-tailed E-selectin with phospholipase C andcollecting the resulting solution, was diluted in a solution composed ofRPMI, HEPES, 0.1% BSA, and 0.1% sodium azide). All compounds tested wereadded prior to the addition Of the labeled HL60 cells and allowed toincubate at 4° C. for one hour.

After the test compound incubation period, 100 μL of labeled HL60 cellswere added to each well, and the microtiter plate was incubated at 4° C.for 30 minutes. Then, the wells were washed three times with RPMI 1640plus 0.1% FCS using a multichannel pipettor. After the wells werewashed, 100 μL of 0.1 N NaOH were added to each well, and the microtiterplate was incubated at room temperature for 10 minutes. After thisincubation, aliquots were removed from each well and placed inscintillation vials; scintillation fluid was added to each vial; and theradioactivity in each aliquot was quantitated by liquid scintillationcounting.

The compounds tested were as follows: (1) "BBA-2" is an E-selectinblocking antibody purchased from R&D Systems, 614 McKinley Place NE,Minneapolis, Minn. 55413 as product code BBG57 and used at aconcentration of about 200 nM; (2) "anti 176" is an antibody that doesnot bind to the E-selectin used at a concentration of about 200 nM as anegative control; (3) "AFFY 4" is peptide HITWDQLWNVMN (SEQ ID NO 23),with a free amino and a free carboxy terminus, used at a concentrationof 200 μM after dissolving in DMSO and diluting in PBS/DMSO to a finalconcentration of 5% DMSO; (4) "AFFY 1" is an unrelated peptide used at aconcentration of about 200 μM as a negative control; and (5) a "SLe^(x)" derivative, monofucosyl monosialyllactose at a concentration of about200 μM. The results of the assay are presented in FIG. 1, which reflectsHL60 cell binding as a percent of the binding observed in the absence ofany added compound.

As shown in FIG. 1, the peptide HITWDQLWNVMN (SEQ ID NO 23) blocked over80% of the cell binding that occurred in the absence of added compound.This result compares favorably with the blocking observed in thepresence of BBA-2 (about 65%). The negative controls (anti 176 andAFFY 1) showed no significant blocking of HL60 binding. Although SLe^(x)did not exhibit significant blocking of cell adhesion at theconcentration used in this assay, subsequent experiments showed thatSLe^(x) does block cell adhesion if used at a concentration of 500 to1000 μM as illustrated in FIG. 2B.

The concentration of any peptide or peptide mimetic required to reduceby 50% the binding of HL60 cells to E-selectin as per the assay ofExample 1 is determined by the following procedure. In this procedurethe IC₅₀ -HL60 of a specific peptide, HITWDQLWNVMN (SEQ ID NO 23), wasdetermined. It being understood that the procedure employed can be usedto the determine the IC₅₀ -HL60 of other peptides and peptide mimetics.

The peptide HITWDQLWNVMN (SEQ ID NO 23) was tested in the cell adhesionassay at varying concentrations (from 0.5 to 100 μM) along with anegative control (the AFFY 1 peptide). The results of this assay,plotted as cpm observed against peptide concentration, are shown in FIG.2A. From this figure, the concentration of the peptide HITWDQLWNVMN (SEQID NO 23) required to inhibit binding of HL60 cells by about 50% isdetermined to be about 50 μM and this value is taken as the IC₅₀ -HL60value for this peptide.

FIG. 4 graphically illustrates the IC₅₀ -HL60 for a second peptideDITWDQLWDLMK (SEQ ID NO 3) which has an IC₅₀ -HL60 of less than about 10μM.

The above results demonstrate that this simple in vitro assay provides afacile means to determine the binding affinity of a candidate peptide orpeptide mimetic to E-selectin as measured by its IC₅₀ -HL60.

FIGS. 3A and 3B illustrate two assay formats for peptides of theinvention the second of which is exemplified in detail in Example 2which follows.

EXAMPLE 2 DETERMINATION OF THE IC₅₀ -STANDARD FOR DIFFERENT PEPTIDES ANDPEPTIDE MIMETICS

This example measures the concentration of peptide or peptide mimeticrequired to inhibit 50% of the binding of a radiolabeled standard toE-selectin and the resulting IC₅₀ value is referred to as the "IC₅₀-standard". In this example, the standard corresponds to the syntheticpeptide LRRASLGDGDITWDQLWDLMK (SEQ ID NO 114) which comprises a knowninitial seven amino acid group which is a substrate for protein kinase Afollowed by a 2 amino acid spacer and a 12 amino acid sequence whichcorresponds to the peptide identified by SEQ ID NO 3 which has an IC₅₀-HL60 of 10 μM. The peptide is radiolabeled by phosphorylation usingeither ³³ p or ³³ P phosphate by conventional methods via the proteinkinase A. The example is a competitive assay in which 6.5 picograms at1400 Ci/mmol (65000 cpm) of radiolabelled standard (1400 Ci/mmol) iscombined with a titered amount of candidate peptide or peptide mimeticin a constant volume of 30 μL. The combined solution is then added toE-selectin immobilized onto a solid surface in the manner describedabove. The resulting mixture is then incubated for about 120 minutes topermit equilibrium binding to be achieved. Afterwards, unbound materialsare separated from bound materials by conventional methods (e.g.,washing) and the amount of the candidate peptide or peptide mimeticbound to E-selectin is determined by scintillation counting. The assayis repeated over several concentrations of candidate peptide or peptidemimetic and the results are entered into a graph whose y axis is theamount of radiolabeled standard bound to E-selectin (measured in cpm)and the x axis is the amount of candidate peptide or peptide mimetic.The results are graphically extrapolated to determine the concentrationat which the peptide or peptide mimetic will reduce by 50% the amount ofradiolabeled standard bound to E-selectin. This concentration is takenas the IC₅₀ -standard for that peptide or peptide mimetic.

Additional peptides were prepared synthetically as free peptides andassayed in the manner described above to determine their IC₅₀ -standardvalues. The results of this assay are shown in the following table:

    ______________________________________                                                                   IC.sub.50 -                                                                   STAN-                                              PEPTIDE SEQUENCE           DARD                                               ______________________________________                                        DITWDQLWDLMK (SEQ ID NO 3) 3 nM                                               DGDITWDQLWDLMK (SEQ ID NO 4)                                                                             4 nM                                               DYTWFELWDMMQ (SEQ ID NO 5) 6 nM                                               DITWDELWKIMN (SEQ ID NO 6) 7 nM                                               QITWAQLWNMMK (SEQ ID NO 7) 12 nM                                              DYSWHDLWEMMS (SEQ ID NO 8) 16 nM                                              DITWDQLWDLN1eK (SEQ ID NO 9)                                                                             23 nM                                              HITWDQLWRIMT (SEQ ID NO 10)                                                                              28 nM                                              RNMSWLELWEHMK (SEQ ID NO 102)                                                                            35 nM                                              DGDITWDQLWDLNleK (SEQ ID NO 113)                                                                         42 nM                                              QITWDQLWDLNleK (SEQ ID NO 115)                                                                           65 nM                                              AEKWDQLWHVMNPAESQ (SEQ ID NO 116)                                                                        65 nM                                              DITWAQLWNNleNleN (SEQ ID No 117)                                                                         425 nM                                             d-DITWDQLWDLMK             25 nM                                              Dd-ITWDQLWDLMK             27 nM                                              DId-TWDQLWDLMK             2700 nM                                            DITWd-DQLWDLMK             34 nM                                              DITWDQLWDd-LMK             33000 nM                                           DITWDQLWDLMd-K             1000 nM                                            DITWDQLWDLMK-CONH.sub.2 (SEQ ID NO 166)                                                                  22 nM                                              ITWDQLWDLMK (SEQ ID N0 160)                                                                              11 nm                                              TWDQLWDLMK (SEQ ID NO 162) 12 μM                                           ______________________________________                                    

As shown above, a number of the peptides have IC₅₀ -standard values ofabout 10 nanomolar (nM) or less.

EXAMPLE 3 BINDING OF PEPTIDES TO SELECTINS OTHER THAN E-SELECTIN

This example demonstrates that certain of the peptides of this inventionalso have binding affinity for selectins other than E-selectin includingthe P-selectin and the L-selectin. In this example, the bindingspecificity of certain peptides on phage for the E-selectin, theP-selectin, the L-selectin and NGF R (control) were conducted using anELISA procedure to determine if the peptides bind specifically to theseselectins. This assay was carried out similarly to the procedures usedin the affinity enrichment process described above, except that afterremoving unbound phage from the wells, the wells were treated withrabbit anti-phage antibody, then with alkaline phosphatase(AP)-conjugated goat anti-rabbit immunoglobulin antibody, or withanti-LacI-AP, as shown in FIG. 3, part A, and then the amount ofalkaline phosphatase in each well was determined measuring the opticaldensity at 405 nm. The amount of alkaline phosphatase in each well isdetermined by incubation of a chromogenic enzyme substrate with thematerial left in the well. If cleaved by the enzyme, a color isproduced. If no enzyme is present, no color appears; this is interpretedto mean that no goat anti-rabbit Ig/AP was bound.

The results of this analysis are set forth in FIG. 5 wherein peptidesfrom families 1 and 2 clearly show binding affinity for each of theselectins as compared to control.

EXAMPLE 4 BINDING OF PEPTIDES CONTAINING SYNTHETIC AMINO ACIDS TOE-SELECTIN

This example demonstrates that peptide containing synthetic amino acidscan bind to the E-selectin. Specifically, in this example, the followingsynthetic amino acids were employed: ##STR5## and 2-amino isobutyricacid (Aib).

The above amino acids, except M(OCH₃), are readily available materials.The synthesis of M(OCH₃) was conducted as shown in reaction (1) belowwhich synthesis follows the procedures set forth in the art for thetritylation step, methylation and detritylation step⁴³,44 : ##STR6##

Specifically, to a stirred suspension of L-homoserine (8 grams, 67 mmol)in methylene chloride (120 mL) was added (CH₃)₃ SiCl (30 mL, 234 mmol),and the mixture was refluxed for 30 minutes. It was then allowed toreach room temperature, treated with a solution of triethylamine (9.34mL, mmol) in 120 mL of methylene chloride and refluxed for 45 minutes.The reaction mixture, at 0° C., was treated dropwise with anhydrousmethanol (4 mL) in methylene chloride (250 mL) and allowed to attainroom temperature. Then triethylamine (9.34 mL, 67 mmol) was addedfollowed by the addition of trityl chloride (Trt-Cl) (18.8 grams, 67mmol) over a 15 minute period. The reaction was stirred overnight.Evaporation under reduced pressure left a residue, which was partitionedbetween ethyl acetate (300 mL) and a 5% precooled aqueous solution ofcitric acid (200 mL). The organic layer was collected, washed with 1 NNaOH (150 mL) and water (150 mL). The combined aqueous layers werewashed with ether (50 mL), cooled to 0° C., and neutralized with glacialacetic acid. The precipitated product was extracted with ethyl acetate(2×200 mL), and the combined organic layers were washed twice withsaturated NaCl and dried (Na₂ SO₃). Evaporation of the solvents in vacuogave compound 1 (18 grams, 75% yield) as a white solid.

To a suspension of sodium hydride (11.9 grams, 80% in oil, 497 mmol) andimidazole (0.639 grams, 9.4 mmol) in tetrahydrofuran (350 mL), asolution of compound 1 (17.3 grams, 47 mmol) in tetrahydrofuran (120 mL)was added with stirring at -15° C. over 15 minutes. After 1 hour at thattemperature, methyl iodide (6.23 mL, 100 mmol) was added and the mixturewas stirred at -5° C. for 10 hours. The resulting mixture wassubsequently diluted, at 0° C. with water (300 mL) and extracted withether (2×100 mL). The cooled aqueous layer was then neutralized withglacial acetic acid and extracted with ethyl acetate. Organic layerswere combined, washed with 5% citric acid and brine, dried (Na₂ SO₃),and concentrated in vacuo. The residue was triturated with hexane togive compound 2 (15 grams, 85% yield) as a white powder.

A solution of compound Z (15 grams, 40 mmol) in ethanol (700 mL) wastreated with acetic acid (71 mL) for 40 hours. The solvents wereevaporated and the residue was triturated with hexane to give compound 3(5 g, 94% yield) as a white crystalline powder.

By following the procedures set forth herein, the methoxy group ofM(OCH₃), as described above, can be replaced by a higher homolog thereofsuch as an alkoxy group of from 2 to 10 carbon atoms.

When compound 2 is employed in the peptide synthesis, it isconventionally converted to the F-moc amino protected derivative. Suchconversion is typically achieved as follows:

A solution of NaHCO₃ (8.4 g, 100 mmol) in water (200 mL) was added to asuspension of compound 3 (5 grams, 37 mmol) in dioxane (200 mL) at 0° C.After 10 minutes of stirring, F-moc succinimide (14.8 grams, 44 mmol)was added slowly to the resulting solution and stirring was continueduntil completion of the reaction (24 hours). The cooled reaction mixturewas diluted with water and extracted with ether (3×50 mL). The aqueouslayer was acidified with acetic acid and extracted with ethyl acetate(3×150 mL). The organic layer was washed with brine, concentrated togive a yellowish precipitate that was triturated with ether to givecompound 4 (6 g, 46% yield) as a white powder.

Each of the above synthetic amino acids as well as 2-amino isobutryicacid were N-protected and the carboxyl group activated by conventionalmethods and then incorporated into several peptide structures. Theresulting peptides so prepared were derivatives of the peptideDITWDQLWDLMK (SEQ ID NO 3) and included the following:

DITWDQLWDLMK-NH₂ (SEQ ID NO 167)

DITNal(1)DQLWDLMK-NH₂ (SEQ ID NO 118)

DITWDQLNal (1) DLMK-NH₂ (SEQ ID NO 119)

DITNal(2)DQLWDLMK-NH₂ (SEQ ID NO 120)

DITWDQLNal (2) DLMK-NH₂ (SEQ ID NO 121)

DITChaDQLWDLMK-NH₂ (SEQ ID NO 122)

DITWDQLChaDLMK-NH₂ (SEQ ID NO 123)

DiTWDQLWDLM(OCH₃)K-NH₂ (SEQ ID NO 124)

DiTWDQLWDLM(SOCH₃)K-NH₂ (SEQ ID NO 125)

DiTWDQLWDLM(SO₂ CH₃)K-NH₂ (SEQ ID NO 126)

DITWDQLW-Aib-LMK-NH₂ (SEQ ID NO 127)

DITWDQLW-Aib-LMK-OH (SEQ ID NO 128)

DITW-Aib-QLWKLMK-OH (SEQ ID NO 129)

DITW-Aib-QLWDLMK-NH₂ (SEQ ID NO 130)

DITW-Aib-QLW-Aib-LMK-NH₂ (SEQ ID NO 131)

DITW-Aib-QLWDLMK-OH (SEQ ID NO 132)

DITW-Aib-QLW-Aib-LMK-OH (SEQ ID NO 133)

AITWDQLWDLNleK-OH (SEQ ID NO 134)

DATWDQLWDLNleK-OH (SEQ ID NO 135)

DITADQLWDLNleK-OH (SEQ ID NO 136)

DITWAQLWDLNleK-OH (SEQ ID NO 137)

DITWDALWDLNleK-OH (SEQ ID NO 138)

DITWDQAWDLNleK-OH (SEQ ID NO 139)

DITWDQLADLNleK-OH (SEQ ID NO 140)

DITWDQLWALNleK-OH (SEQ ID NO 141)

DITWDQLWDANleK-OH (SEQ ID NO 142)

DITWDQLWDLAK-OH (SEQ ID NO 143)

DITWDQLWDLNleA-OH (SEQ ID NO 144)

DITNal(1)DQLNal(1)DLMK-NH₂ (SEQ ID NO 145)

DITWAQLNal(1)DLMK-NH₂ (SEQ ID NO 146)

DITNal(1)AQLNal(1)DLMK-NH₂ (SEQ ID NO 147 )

DITNal(1)AQLNal(1)DLM (OCH₃)K-NH₂ (SEQ ID NO 148 )

DiTWAQLWDLM(SO₂ CH₃)K-NH₂ (SEQ ID NO 149)

DiTWAQLNal(1)DLM(SO₂ CH₃)K-NH₂ (SEQ ID NO 150)

DiTNal(1)AQLWDLM(SO₂ CH₃)K-NH₂ (SEQ ID NO 151)

DITWAQLWDLM(OCH₃)K-NH₂ (SEQ ID NO 152)

DITNal(1)AQLWDLMK-NH₂ (SEQ ID NO 153)

DiTNal(1)DQLWDLM(SO₂ CH₃)K-NH₂ (SEQ ID NO 154)

DiTWDQLNal(1)DLM(SO₂ CH₃)K-NH₂ (SEQ ID NO 155)

DiTWAQLWDLMK-NH₂ (SEQ ID NO 156)

DITWAibQLWDLMK-NH₂ (SEQ ID NO 157)

Each of these peptides were assayed to determine their IC₅₀ -standard inthe manner described above in Example 2. The results of this assayindicated that each of the above peptides had an IC₅₀ -standard of lessthan 1 mM indicating that peptides having one or more synthetic aminoacids can nevertheless bind to E-selectin.

EXAMPLE 5 BINDING OF PEPTIDES MIMETICS TO E-SELECTIN

The following example demonstrates that peptide mimetics can also bindto E-selectin. Specifically, in this example, peptide mimetics having aC-terminal --C(O)NH₂ substituent were prepared by conventional solidphase synthetic methods. Other peptide mimetics prepared includedN-terminal modifications such as acetylation (Ac), formation of asuccinimide group at the N-terminus, or the formation of a CBZ group atthe N-terminus via conventional methods described above. Examples ofsuch peptide mimetics include the following:

Ac-DITWDQLWKLMK-OH (SEQ ID NO 168)

Ac-DITWDQLWDL-Nle-K-NH₂ (SEQ ID NO 158)

Succinimide-ITWDQLWDLMK-OH (SEQ ID NO 161)

Cbz-TWDQLWDLMK-OH (SEQ ID NO 163)

Succ-ITWDQLWDLMK-NH₂ (SEQ ID NO 159)

Cbz-DITWDQLWDLMK-NH₂ (SEQ ID NO 160)

Ac-DITWDQLWDLMK-NH₂ (SEQ ID NO 165)

(Ac=acetyl; Succ=succinimidyl)

Each of these peptide mimetics were assayed to determine their IC₅₀-standard in the manner described above in Example 2. The results ofthis assay indicated that each of the above peptide mimetics had an IC₅₀-standard of less than 1 mM indicating that peptide mimetics includingthose having more than one modification can nevertheless bind toE-selectin. The above results further indicate that the amino acidM(OCH₃) is a suitable replacement for methionine in the above peptidesand, accordingly, is contemplated as a suitable for replacement inpeptides containing methionine. In this embodiment, the presentinvention is directed to a method for creating a peptide mimetic whichmethod comprises replacing at least one methionine amino acid groups ina peptide with M(OCH₃).

Although the foregoing invention has been described in some detail byway of illustration and example for purposes of clarity andunderstanding, it will be apparent that certain changes andmodifications may be practiced within the scope of the appended claims.

    __________________________________________________________________________    SEQUENCE LISTING                                                              (1) GENERAL INFORMATION:                                                      (iii) NUMBER OF SEQUENCES: 169                                                (2) INFORMATION FOR SEQ ID NO:1:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 8 amino acids                                                     (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(2, 3, 6, 7)                                              (D) OTHER INFORMATION: /note= "Xaa is any amino acid."                        (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1:                                       TrpXaaXaaLeuTrpXaaXaaMet                                                      15                                                                            (2) INFORMATION FOR SEQ ID NO:2:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(1)                                                       (D) OTHER INFORMATION: /note= "Xaa is His, Glu or Asp."                       (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(2)                                                       (D) OTHER INFORMATION: /note= "Xaa is Ile, Met or                             Norleucine."                                                                  (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(3)                                                       (D) OTHER INFORMATION: /note= "Xaa is Thr or Ser."                            (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(5)                                                       (D) OTHER INFORMATION: /note= "Xaa is Asp, Ala, Glu or                        Leu."                                                                         (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(10)                                                      (D) OTHER INFORMATION: /note= "Xaa is Leu, Met, Val or                        Ile."                                                                         (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(11)                                                      (D) OTHER INFORMATION: /note= "Xaa is Met, Norleucine,                        o-methyl methionine or met-sulfone."                                          (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "Xaa is Asn, Ser, Lys or                        Gln."                                                                         (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2:                                       XaaXaaXaaTrpXaaGlxLeuTrpAsxXaaXaaXaa                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:3:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3:                                       AspIleThrTrpAspGlnLeuTrpAspLeuMetLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:4:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4:                                       AspGlyAspIleThrTrpAspGlnLeuTrpAspLeuMetLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:5:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:5:                                       AspTyrThrTrpPheGluLeuTrpAspMetMetGln                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:6:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6:                                       AspIleThrTrpAspGluLeuTrpLysIleMetAsn                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:7:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7:                                       GlnIleThrTrpAlaGlnLeuTrpAsnMetMetLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:8:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8:                                       AspTyrSerTrpHisAspLeuTrpGluMetMetSer                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:9:                                              (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(11)                                                      (D) OTHER INFORMATION: /note= "Xaa is Norleucine."                            (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9:                                       AspIleThrTrpAspGlnLeuTrpAspLeuXaaLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:10:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:10:                                      HisIleThrTrpAspGlnLeuTrpArgIleMetThr                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:11:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 10 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:11:                                      TyrAspAspValCysCysGluLeuLeuPhe                                                1510                                                                          (2) INFORMATION FOR SEQ ID NO:12:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 10 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:12:                                      AspLeuProGlnTrpTyrThrGluTrpCys                                                1510                                                                          (2) INFORMATION FOR SEQ ID NO:13:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 10 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:13:                                      GluAsnSerHisTrpCysThrCysProCys                                                1510                                                                          (2) INFORMATION FOR SEQ ID NO:14:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 10 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:14:                                      AspIleGluGlnAspTrpValThrTrpMet                                                1510                                                                          (2) INFORMATION FOR SEQ ID NO:15:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 10 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:15:                                      AsnGluTrpCysValValProCysArgLeu                                                1510                                                                          (2) INFORMATION FOR SEQ ID NO:16:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 10 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:16:                                      AspIleTrpGlnAspTrpValArgTrpMet                                                1510                                                                          (2) INFORMATION FOR SEQ ID NO:17:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 10 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:17:                                      AspLeuTrpGlnAspTrpValThrTrpMet                                                1510                                                                          (2) INFORMATION FOR SEQ ID NO:18:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 10 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:18:                                      AspLeuTrpGlnAspTrpValHisTrpMet                                                1510                                                                          (2) INFORMATION FOR SEQ ID NO:19:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 10 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:19:                                      AspIleTrpGlnAspTrpValThrTrpMet                                                1510                                                                          (2) INFORMATION FOR SEQ ID NO:20:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 10 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:20:                                      AspIleTrpGlnAspTrpValLysTrpMet                                                1510                                                                          (2) INFORMATION FOR SEQ ID NO:21:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 10 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:21:                                      AspIleTrpGluAspTrpValArgTrpMet                                                1510                                                                          (2) INFORMATION FOR SEQ ID NO:22:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 10 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:22:                                      AspIleTrpGlnAspTrpIleThrTrpMet                                                1510                                                                          (2) INFORMATION FOR SEQ ID NO:23:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:23:                                      HisIleThrTrpAspGlnLeuTrpAsnValMetAsn                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:24:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:24:                                      HisIleThrTrpAspGlnXaaXaaXaaXaaXaaXaa                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:25:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:25:                                      XaaXaaThrTrpAspGlnLeuTrpXaaXaaXaaXaa                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:26:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:26:                                      XaaXaaXaaXaaAspGlnLeuTrpAsnValXaaXaa                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:27:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:27:                                      XaaXaaXaaXaaXaaXaaLeuTrpAsnValMetAsn                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:28:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:28:                                      XaaXaaXaaXaaHisIleThrTrpAspGlnLeuTrpAsnValMetAsn                              151015                                                                        XaaXaaXaaXaa                                                                  20                                                                            (2) INFORMATION FOR SEQ ID NO:29:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:29:                                      XaaXaaXaaTrpXaaXaaLeuTrpXaaXaaMetXaa                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:30:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 18 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:30:                                      XaaXaaXaaXaaTrpXaaXaaLeuTrpXaaXaaMetXaaXaaXaaXaa                              151015                                                                        XaaXaa                                                                        (2) INFORMATION FOR SEQ ID NO:31:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:31:                                      AlaIleThrTrpAspGlnLeuTrpLeuLeuMetSer                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:32:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:32:                                      GluLeuThrTrpAspGlnLeuTrpValLeuMetSer                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:33:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:33:                                      AspValThrTrpAspGlnLeuTrpGluLeuMetThr                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:34:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:34:                                      GluValThrTrpAspGlnLeuTrpValMetMetGln                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:35:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:35:                                      AsnLeuThrTrpAspGlnLeuTrpValLeuMetSer                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:36:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:36:                                      GluMetSerTrpLeuGluLeuTrpAsnValMetAsn                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:37:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:37:                                      ThrIleThrTrpAspGlnLeuTrpGlnMetMetSer                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:38:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:38:                                      GluLeuSerTrpAspGlnLeuTrpAsnValMetAsn                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:39:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:39:                                      GluMetThrTrpGlnGluLeuTrpAsnValMetAsn                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:40:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:40:                                      GluMetThrTrpThrGluLeuTrpAsnValMetAsn                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:41:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:41:                                      AspMetThrTrpSerGlnLeuTrpAsnValMetAsn                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:42:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:42:                                      GluMetThrTrpLeuGlyLeuTrpAsnValMetAsn                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:43:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:43:                                      GlnIleThrTrpMetGluLeuTrpAsnLeuMetAsn                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:44:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:44:                                      GluIleThrTrpAspGlnLeuTrpGluValMetAsn                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:45:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:45:                                      GluIleThrTrpAspGlnLeuTrpAspValMetAsn                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:46:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:46:                                      AspIleSerTrpAspGlnLeuTrpAsnValMetAsn                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:47:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:47:                                      GlnIleThrTrpAspGlnLeuTrpAspLeuMetLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:48:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:48:                                      GluMetThrTrpAspGlnLeuTrpAsnValMetAsn                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:49:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:49:                                      AspIleThrTrpAspGlnLeuTrpAsnMetMetAsp                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:50:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:50:                                      AspIleThrTrpAsnMetLeuTrpAsnMetMetGln                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:51:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:51:                                      AspIleSerTrpAspAspLeuTrpIleMetMetAsn                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:52:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:52:                                      AspIleThrTrpHisGlnLeuTrpAsnLeuMetAsn                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:53:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:53:                                      GluIleSerTrpGluGlnLeuTrpThrMetMetAsn                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:54:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:54:                                      AspIleThrTrpGluGlnLeuTrpAsnMetMetAsn                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:55:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:55:                                      GluIleThrTrpAspGlnLeuTrpThrLeuMetThr                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:56:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:56:                                      AspIleThrTrpHisGlnLeuTrpAsnLeuMetAsn                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:57:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:57:                                      AspMetThrTrpAspGlnLeuTrpIleValMetAsn                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:58:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:58:                                      AspIleThrTrpGluGlnLeuTrpAsnLeuMetAsn                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:59:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:59:                                      GlnIleThrTrpTyrGlnLeuTrpAsnMetMetAsn                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:60:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:60:                                      HisIleSerTrpHisGluLeuTrpAsnLeuMetGln                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:61:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:61:                                      TyrIleThrTrpGluGlnLeuTrpThrMetMetAsn                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:62:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:62:                                      HisIleThrTrpAspGlnLeuTrpAspLeuMetGln                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:63:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:63:                                      GlnIleThrTrpAspGlnLeuTrpAspLeuMetTyr                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:64:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:64:                                      GlnIleThrTrpAspGlnLeuTrpAsnMetMetIle                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:65:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:65:                                      TyrIleThrTrpGluGlnLeuTrpAsnMetMetAsn                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:66:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:66:                                      HisIleThrTrpAspGlnLeuTrpAspIleMetSer                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:67:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:67:                                      HisIleThrTrpAspGlnLeuTrpGluIleMetSer                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:68:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:68:                                      HisIleThrTrpAspGlnLeuTrpAlaLeuMetThr                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:69:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:69:                                      HisIleThrTrpAspGlnLeuTrpSerLeuMetSer                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:70:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:70:                                      HisIleThrTrpAspGlnLeuTrpLeuMetMetSer                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:71:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:71:                                      HisIleThrTrpAspGlnLeuTrpAspLeuMetGln                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:72:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:72:                                      HisIleThrTrpAspGlnLeuTrpTrpIleMetAla                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:73:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:73:                                      HisIleThrTrpAspGlnLeuTrpLeuLeuMetAla                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:74:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:74:                                      HisIleThrTrpAspGlnLeuTrpMetLeuMetAla                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:75:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:75:                                      GlySerAspSerHisIleThrTrpAspGluLeuTrpAsnLeuMetAsn                              151015                                                                        ProValLeuAla                                                                  20                                                                            (2) INFORMATION FOR SEQ ID NO:76:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:76:                                      AsnTrpLeuAspAspIleThrTrpAspGluLeuTrpLysIleMetAsn                              151015                                                                        ProSerThrAla                                                                  20                                                                            (2) INFORMATION FOR SEQ ID NO:77:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:77:                                      GluThrAspAspHisIleThrTrpAspGlnLeuTrpArgIleMetThr                              151015                                                                        AlaThrMetAla                                                                  20                                                                            (2) INFORMATION FOR SEQ ID NO:78:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:78:                                      TrpThrAspThrHisIleThrTrpAspGlnLeuTrpHisPheMetAsn                              151015                                                                        MetGlyGluGln                                                                  20                                                                            (2) INFORMATION FOR SEQ ID NO:79:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:79:                                      GlyPheGlyGluAlaIleThrTrpAspGlnLeuTrpAspMetMetAsn                              151015                                                                        GlyGluAspAla                                                                  20                                                                            (2) INFORMATION FOR SEQ ID NO:80:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:80:                                      AsnValAlaGluGlnIleThrTrpAspGlnLeuTrpAsnLeuMetSer                              151015                                                                        ValGlySerSer                                                                  20                                                                            (2) INFORMATION FOR SEQ ID NO:81:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:81:                                      GlyGlnThrGlyLeuIleThrTrpAspMetLeuTrpAsnLeuMetAsn                              151015                                                                        ProValGlyGlu                                                                  20                                                                            (2) INFORMATION FOR SEQ ID NO:82:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:82:                                      GlyThrGlyAspHisIleThrTrpAspGlnLeuTrpAsnLeuMetIle                              151015                                                                        AsnGluLysGly                                                                  20                                                                            (2) INFORMATION FOR SEQ ID NO:83:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:83:                                      GluTyrGlyArgHisIleThrTrpAspGlnLeuTrpGlnLeuMetGln                              151015                                                                        SerAlaThrAla                                                                  20                                                                            (2) INFORMATION FOR SEQ ID NO:84:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:84:                                      MetAsnAsnTrpHisValSerTrpGluGlnLeuTrpAspIleMetAsn                              151015                                                                        GlyProProAsn                                                                  20                                                                            (2) INFORMATION FOR SEQ ID NO:85:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:85:                                      GluSerAlaSerHisIleThrTrpGlyGlnLeuTrpAspLeuMetAsn                              151015                                                                        AlaSerGluVal                                                                  20                                                                            (2) INFORMATION FOR SEQ ID NO:86:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:86:                                      TyrTrpArgGlyAsnIleThrTrpAspGlnLeuTrpAsnIleMetAsn                              151015                                                                        SerGluTyrSer                                                                  20                                                                            (2) INFORMATION FOR SEQ ID NO:87:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:87:                                      AlaGlyAlaSerHisIleThrTrpAlaGlnLeuTrpAsnMetMetAsn                              151015                                                                        GlyAsnGluGly                                                                  20                                                                            (2) INFORMATION FOR SEQ ID NO:88:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:88:                                      GluSerTrpAlaHisIleThrTrpAspGlnLeuTrpAsnLeuMetAsn                              151015                                                                        MetGlyThrGln                                                                  20                                                                            (2) INFORMATION FOR SEQ ID NO:89:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:89:                                      TyrGlyAsnSerAsnIleThrTrpAspGlnLeuTrpSerIleMetAsn                              151015                                                                        ArgGlnThrThr                                                                  20                                                                            (2) INFORMATION FOR SEQ ID NO:90:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:90:                                      AlaHisLeuProHisIleSerTrpAspThrLeuTrpHisIleMetAsn                              151015                                                                        LysGlyGluLys                                                                  20                                                                            (2) INFORMATION FOR SEQ ID NO:91:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:91:                                      GluSerAlaSerHisIleThrTrpGlyGlnLeuTrpAspLeuMetAsn                              151015                                                                        AlaSerGluVal                                                                  20                                                                            (2) INFORMATION FOR SEQ ID NO:92:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:92:                                      MetAsnAsnTrpHisValSerTrpGluGlnLeuTrpAspIleMetAsn                              151015                                                                        GlyProProAsn                                                                  20                                                                            (2) INFORMATION FOR SEQ ID NO:93:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:93:                                      GlyPheGlyGluAlaIleThrTrpAspGlnLeuTrpAspMetMetAsn                              151015                                                                        GlyGluAspAla                                                                  20                                                                            (2) INFORMATION FOR SEQ ID NO:94:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 20 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:94:                                      TrpThrAspThrHisIleThrTrpAspGlnLeuTrpHisPheMetAsn                              151015                                                                        MetGlyGluGln                                                                  20                                                                            (2) INFORMATION FOR SEQ ID NO:95:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:95:                                      GluMetThrTrpAlaGluLeuTrpThrLeuMetGlu                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:96:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:96:                                      AspIleSerTrpArgGlnLeuTrpAspIleMetAsn                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:97:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:97:                                      GluIleSerTrpLeuGlyLeuTrpAspIleMetAsn                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:98:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:98:                                      AspMetThrTrpHisAspLeuTrpThrLeuMetSer                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:99:                                             (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:99:                                      ArgGlyValTrpGlyGlyLeuTrpSerMetThrTrp                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:100:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:100:                                     GluMetThrTrpGlnGlnLeuTrpValValMetGln                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:101:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 18 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:101:                                     AlaGluTrpThrTrpAspGlnLeuTrpHisValMetAsnProAlaGlu                              151015                                                                        SerGln                                                                        (2) INFORMATION FOR SEQ ID NO:102:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 13 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:102:                                     ArgAsnMetSerTrpLeuGluLeuTrpGluHisMetLys                                       1510                                                                          (2) INFORMATION FOR SEQ ID NO:103:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 18 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:103:                                     SerGlnValThrTrpAsnAspLeuTrpSerValMetAsnProGluVal                              151015                                                                        ValAsn                                                                        (2) INFORMATION FOR SEQ ID NO:104:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:104:                                     HisArgAlaGluTrpLeuAlaLeuTrpGluGlnMetSerPro                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:105:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:105:                                     TyrLysLysGluTrpLeuGluLeuTrpHisGlnMetGlnAla                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:106:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 13 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:106:                                     ArgSerLeuSerTrpLeuGlnLeuTrpAspGlnMetLys                                       1510                                                                          (2) INFORMATION FOR SEQ ID NO:107:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 13 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:107:                                     LysGluGlnGlnTrpArgAsnLeuTrpLysMetMetSer                                       1510                                                                          (2) INFORMATION FOR SEQ ID NO:108:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 16 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:108:                                     LysLysGluAspTrpLeuAlaLeuTrpArgIleMetSerValProAsp                              151015                                                                        (2) INFORMATION FOR SEQ ID NO:109:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 13 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:109:                                     ArgAsnMetSerTrpLeuGluLeuTrpGluHisMetLys                                       1510                                                                          (2) INFORMATION FOR SEQ ID NO:110:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 15 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:110:                                     GlyArgProThrTrpAsnGluLeuTrpAspMetMetGlnAlaPro                                 151015                                                                        (2) INFORMATION FOR SEQ ID NO:111:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 13 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:111:                                     LysArgLysGlnTrpIleGluLeuTrpAsnIleMetSer                                       1510                                                                          (2) INFORMATION FOR SEQ ID NO:112:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:112:                                     LysThrSerGluTrpAsnAsnLeuTrpLysLeuMetSerGln                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:113:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 14 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(13)                                                      (D) OTHER INFORMATION: /note= "Xaa is Norleucine."                            (xi) SEQUENCE DESCRIPTION: SEQ ID NO:113:                                     AspGlyAspIleThrTrpAspGlnLeuTrpAspLeuXaaLys                                    1510                                                                          (2) INFORMATION FOR SEQ ID NO:114:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 21 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:114:                                     LeuArgArgAlaSerLeuGlyAspGlyAspIleThrTrpAspGlnLeu                              151015                                                                        TrpAspLeuMetLys                                                               20                                                                            (2) INFORMATION FOR SEQ ID NO:115:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(11)                                                      (D) OTHER INFORMATION: /note= "Xaa is Norleucine."                            (xi) SEQUENCE DESCRIPTION: SEQ ID NO:115:                                     GlnIleThrTrpAspGlnLeuTrpAspLeuXaaLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:116:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 17 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:116:                                     AlaGluLysTrpAspGlnLeuTrpHisValMetAsnProAlaGluSer                              151015                                                                        Gln                                                                           (2) INFORMATION FOR SEQ ID NO:117:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(10,11)                                                   (D) OTHER INFORMATION: /note= "Xaa is Norleucine."                            (xi) SEQUENCE DESCRIPTION: SEQ ID NO:117:                                     AspIleThrTrpAlaGlnLeuTrpAsnXaaXaaAsn                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:118:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(4)                                                       (D) OTHER INFORMATION: /note= "Xaa is Nal(1)."                                (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is amidated."                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:118:                                     AspIleThrXaaAspGlnLeuTrpAspLeuMetLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:119:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(8)                                                       (D) OTHER INFORMATION: /note= "Xaa is Nal(1)."                                (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is amidated."                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:119:                                     AspIleThrTrpAspGlnLeuXaaAspLeuMetLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:120:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(4)                                                       (D) OTHER INFORMATION: /note= "Xaa is Nal(2)."                                (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is amidated."                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:120:                                     AspIleThrXaaAspGlnLeuTrpAspLeuMetLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:121:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(8)                                                       (D) OTHER INFORMATION: /note= "Xaa is Nal(2)."                                (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is amidated."                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:121:                                     AspIleThrTrpAspGlnLeuXaaAspLeuMetLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:122:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(4)                                                       (D) OTHER INFORMATION: /note= "Xaa is Cha."                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is amidated."                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:122:                                     AspIleThrXaaAspGlnLeuTrpAspLeuMetLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:123:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(8)                                                       (D) OTHER INFORMATION: /note= "Xaa is Cha."                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is amidated."                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:123:                                     AspIleThrTrpAspGlnLeuXaaAspLeuMetLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:124:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(11)                                                      (D) OTHER INFORMATION: /note= "Xaa is M(OCH-3)."                              (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is amidated."                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:124:                                     AspIleThrTrpAspGlnLeuTrpAspLeuXaaLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:125:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(11)                                                      (D) OTHER INFORMATION: /note= "Xaa is M(SOCH-3)."                             (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is amidated."                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:125:                                     AspIleThrTrpAspGlnLeuTrpAspLeuXaaLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:126:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(11)                                                      (D) OTHER INFORMATION: /note= "Xaa is M(SO-2CH-3)."                           (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is amidated."                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:126:                                     AspIleThrTrpAspGlnLeuTrpAspLeuXaaLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:127:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(9)                                                       (D) OTHER INFORMATION: /note= "Xaa is 2-amino isobutyric                      acid."                                                                        (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is amidated."                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:127:                                     AspIleThrTrpAspGlnLeuTrpXaaLeuMetLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:128:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(9)                                                       (D) OTHER INFORMATION: /note= "Xaa is 2-amino isobutyric                      acid."                                                                        (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is                              hydroxylated."                                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:128:                                     AspIleThrTrpAspGlnLeuTrpXaaLeuMetLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:129:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(5)                                                       (D) OTHER INFORMATION: /note= "Xaa is 2-amino isobutyric                      acid."                                                                        (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is                              hydroxylated."                                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:129:                                     AspIleThrTrpXaaGlnLeuTrpLysLeuMetLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:130:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(5)                                                       (D) OTHER INFORMATION: /note= "Xaa is 2-amino isobutyric                      acid."                                                                        (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is amidated."                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:130:                                     AspIleThrTrpXaaGlnLeuTrpAspLeuMetLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:131:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(5)                                                       (D) OTHER INFORMATION: /note= "Xaa is 2-amino isobutyric                      acid."                                                                        (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(9)                                                       (D) OTHER INFORMATION: /note= "Xaa is 2-amino isobutyric                      acid."                                                                        (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is amidated."                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:131:                                     AspIleThrTrpXaaGlnLeuTrpXaaLeuMetLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:132:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(5)                                                       (D) OTHER INFORMATION: /note= "Xaa is 2-amino isobutyric                      acid."                                                                        (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is                              hydroxylated."                                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:132:                                     AspIleThrTrpXaaGlnLeuTrpAspLeuMetLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:133:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(5)                                                       (D) OTHER INFORMATION: /note= "Xaa is 2-amino isobutyric                      acid."                                                                        (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(9)                                                       (D) OTHER INFORMATION: /note= "Xaa is 2-amino isobutyric                      acid."                                                                        (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is                              hydroxylated."                                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:133:                                     AspIleThrTrpXaaGlnLeuTrpXaaLeuMetLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:134:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(11)                                                      (D) OTHER INFORMATION: /note= "Xaa is Norleucine."                            (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is                              hydroxylated."                                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:134:                                     AlaIleThrTrpAspGlnLeuTrpAspLeuXaaLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:135:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(11)                                                      (D) OTHER INFORMATION: /note= "Xaa is Norleucine."                            (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is                              hydroxylated."                                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:135:                                     AspAlaThrTrpAspGlnLeuTrpAspLeuXaaLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:136:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(11)                                                      (D) OTHER INFORMATION: /note= "Xaa is Norleucine."                            (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is                              hydroxylated."                                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:136:                                     AspIleThrAlaAspGlnLeuTrpAspLeuXaaLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:137:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(11)                                                      (D) OTHER INFORMATION: /note= "Xaa is Norleucine."                            (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is                              hydroxylated."                                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:137:                                     AspIleThrTrpAlaGlnLeuTrpAspLeuXaaLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:138:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(11)                                                      (D) OTHER INFORMATION: /note= "Xaa is Norleucine."                            (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is                              hydroxylated."                                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:138:                                     AspIleThrTrpAspAlaLeuTrpAspLeuXaaLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:139:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(11)                                                      (D) OTHER INFORMATION: /note= "Xaa is Norleucine."                            (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is                              hydroxylated."                                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:139:                                     AspIleThrTrpAspGlnAlaTrpAspLeuXaaLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:140:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(11)                                                      (D) OTHER INFORMATION: /note= "Xaa is Norleucine."                            (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is                              hydroxylated."                                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:140:                                     AspIleThrTrpAspGlnLeuAlaAspLeuXaaLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:141:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(11)                                                      (D) OTHER INFORMATION: /note= "Xaa is Norleucine."                            (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is                              hydroxylated."                                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:141:                                     AspIleThrTrpAspGlnLeuTrpAlaLeuXaaLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:142:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(11)                                                      (D) OTHER INFORMATION: /note= "Xaa is Norleucine."                            (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is                              hydroxylated."                                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:142:                                     AspIleThrTrpAspGlnLeuTrpAspAlaXaaLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:143:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is                              hydroxylated."                                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:143:                                     AspIleThrTrpAspGlnLeuTrpAspLeuAlaLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:144:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(11)                                                      (D) OTHER INFORMATION: /note= "Xaa is Norleucine."                            (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Ala is                              hydroxylated."                                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:144:                                     AspIleThrTrpAspGlnLeuTrpAspLeuXaaAla                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:145:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(4)                                                       (D) OTHER INFORMATION: /note= "Xaa is Nal(1)."                                (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(8)                                                       (D) OTHER INFORMATION: /note= "Xaa is Nal(1)."                                (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is amidated."                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:145:                                     AspIleThrXaaAspGlnLeuXaaAspLeuMetLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:146:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(8)                                                       (D) OTHER INFORMATION: /note= "Xaa is Nal(1)."                                (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is amidated."                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:146:                                     AspIleThrTrpAlaGlnLeuXaaAspLeuMetLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:147:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(4)                                                       (D) OTHER INFORMATION: /note= "Xaa is Nal(1)."                                (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(8)                                                       (D) OTHER INFORMATION: /note= "Xaa is Nal(1)."                                (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is amidated."                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:147:                                     AspIleThrXaaAlaGlnLeuXaaAspLeuMetLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:148:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(4)                                                       (D) OTHER INFORMATION: /note= "Xaa is Nal(1)."                                (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(8)                                                       (D) OTHER INFORMATION: /note= "Xaa is Nal(1)."                                (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(11)                                                      (D) OTHER INFORMATION: /note= "Xaa is M(OCH-3)."                              (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is amidated."                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:148:                                     AspIleThrXaaAlaGlnLeuXaaAspLeuXaaLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:149:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(11)                                                      (D) OTHER INFORMATION: /note= "Xaa is M(SO-2CH-3)."                           (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is amidated."                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:149:                                     AspIleThrTrpAlaGlnLeuTrpAspLeuXaaLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:150:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(8)                                                       (D) OTHER INFORMATION: /note= "Xaa is Nal(1)."                                (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(11)                                                      (D) OTHER INFORMATION: /note= "Xaa is M(SO-2CH-3)."                           (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is amidated."                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:150:                                     AspIleThrTrpAlaGlnLeuXaaAspLeuXaaLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:151:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(4)                                                       (D) OTHER INFORMATION: /note= "Xaa is Nal(1)."                                (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(11)                                                      (D) OTHER INFORMATION: /note= "Xaa is M(SO-2CH-3)."                           (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is amidated."                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:151:                                     AspIleThrXaaAlaGlnLeuTrpAspLeuXaaLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:152:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(11)                                                      (D) OTHER INFORMATION: /note= "Xaa is M(OCH-3)."                              (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is amidated."                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:152:                                     AspIleThrTrpAlaGlnLeuTrpAspLeuXaaLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:153:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(4)                                                       (D) OTHER INFORMATION: /note= "Xaa is Nal(1)."                                (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is amidated."                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:153:                                     AspIleThrXaaAlaGlnLeuTrpAspLeuMetLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:154:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(4)                                                       (D) OTHER INFORMATION: /note= "Xaa is Nal(1)."                                (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(11)                                                      (D) OTHER INFORMATION: /note= "Xaa is M(SO-2CH-3)."                           (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is amidated."                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:154:                                     AspIleThrXaaAspGlnLeuTrpAspLeuXaaLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:155:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(8)                                                       (D) OTHER INFORMATION: /note= "Xaa is Nal(1)."                                (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(11)                                                      (D) OTHER INFORMATION: /note= "Xaa is M(SO-2CH-3)."                           (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is amidated."                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:155:                                     AspIleThrTrpAspGlnLeuXaaAspLeuXaaLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:156:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is amidated."                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:156:                                     AspIleThrTrpAlaGlnLeuTrpAspLeuMetLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:157:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(5)                                                       (D) OTHER INFORMATION: /note= "Xaa is 2-amino isobutyric                      acid."                                                                        (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is amidated."                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:157:                                     AspIleThrTrpXaaGlnLeuTrpAspLeuMetLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:158:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(1)                                                       (D) OTHER INFORMATION: /note= "N-terminal Asp is acetylated."                 (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(11)                                                      (D) OTHER INFORMATION: /note= "Xaa is Norleucine."                            (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is amidated."                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:158:                                     AspIleThrTrpAspGlnLeuTrpAspLeuXaaLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:159:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 11 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(1)                                                       (D) OTHER INFORMATION: /note= "N-terminal Ile is                              succinimidylated."                                                            (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(11)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is amidated."                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:159:                                     IleThrTrpAspGlnLeuTrpAspLeuMetLys                                             1510                                                                          (2) INFORMATION FOR SEQ ID NO:160:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 11 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:160:                                     IleThrTrpAspGlnLeuTrpAspLeuMetLys                                             1510                                                                          (2) INFORMATION FOR SEQ ID NO:161:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 11 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(1)                                                       (D) OTHER INFORMATION: /note= "N-terminal Ile is modified with                succinimide."                                                                 (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(11)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is                              hydroxylated."                                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:161:                                     IleThrTrpAspGlnLeuTrpAspLeuMetLys                                             1510                                                                          (2) INFORMATION FOR SEQ ID NO:162:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 10 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:162:                                     ThrTrpAspGlnLeuTrpAspLeuMetLys                                                1510                                                                          (2) INFORMATION FOR SEQ ID NO:163:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 10 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(1)                                                       (D) OTHER INFORMATION: /note= "N-terminal Thr is modified with                benxyloxycarbonyl."                                                           (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(10)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is                              hydroxylated."                                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:163:                                     ThrTrpAspGlnLeuTrpAspLeuMetLys                                                1510                                                                          (2) INFORMATION FOR SEQ ID NO:164:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 10 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(10)                                                      (D) OTHER INFORMATION: /note= "C-terminal Met is amidated."                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:164:                                     AspIleTrpGlnAspTrpValArgTrpMet                                                1510                                                                          (2) INFORMATION FOR SEQ ID NO:165:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(1)                                                       (D) OTHER INFORMATION: /note= "N-terminal Asp is acetylated."                 (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is amidated."                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:165:                                     AspIleThrTrpAspGlnLeuTrpAspLeuMetLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:166:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is modified with                CONH-2."                                                                      (xi) SEQUENCE DESCRIPTION: SEQ ID NO:166:                                     AspIleThrTrpAspGlnLeuTrpAspLeuMetLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:167:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is amidated."                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:167:                                     AspIleThrTrpAspGlnLeuTrpAspLeuMetLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:168:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(1)                                                       (D) OTHER INFORMATION: /note= "N-terminal Asp is acetylated."                 (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is                              hydroxylated."                                                                (xi) SEQUENCE DESCRIPTION: SEQ ID NO:168:                                     AspIleThrTrpAspGlnLeuTrpLysLeuMetLys                                          1510                                                                          (2) INFORMATION FOR SEQ ID NO:169:                                            (i) SEQUENCE CHARACTERISTICS:                                                 (A) LENGTH: 12 amino acids                                                    (B) TYPE: amino acid                                                          (C) STRANDEDNESS: single                                                      (D) TOPOLOGY: linear                                                          (ii) MOLECULE TYPE: peptide                                                   (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(1)                                                       (D) OTHER INFORMATION: /note= "N-terminal Asp is modified with                benzyloxycarbonyl."                                                           (ix) FEATURE:                                                                 (A) NAME/KEY: Region                                                          (B) LOCATION: one-of(12)                                                      (D) OTHER INFORMATION: /note= "C-terminal Lys is amidated."                   (xi) SEQUENCE DESCRIPTION: SEQ ID NO:169:                                     AspIleThrTrpAspGlnLeuTrpAspLeuMetLys                                          1510                                                                          __________________________________________________________________________

What is claimed is:
 1. A method for treating disease and diseasesymptoms which are mediated by the presence of ELAM-1 which methodcomprises the administration to a patient of an amount sufficient toreduce inflammation of a pharmaceutical composition comprising a peptideand/or peptide mimetic and a pharmaceutically acceptable carrier,wherein said peptide or peptide mimetic has a core sequence comprisingWXXLWXXM (SEQ ID NO:1) where each amino acid is indicated by standardone letter abbreviations, and X is any amino acid.
 2. The method ofclaim 1, wherein said peptide or peptide mimetic is 9 to 12 amino acidresidues in length and comprises the sequence X₁ X₂ X₃ WX₄ X₅ LWX₆ X₇ X₈X₉ (SEQ ID NO:2) wherein each residue can be independently selected asfollows: X₁ is H, E, or D; X₂ is I, M, or Nle; X₃ is T or S; X₄ is D, E,or L; X₅ is Q or E; X₆ is N or D; X₇ is L, M, V, or I; X₈ is M or Nle;and X₉ is N, S or Q.
 3. The method of claim 2, wherein said peptide isselected from the group consisting of a peptide comprising thesequence:DITWDQLWDLMK (SEQ ID NO:3) DGDITWDQLWDLMK (SEQ ID NO:4);DITWDQLWDLNleK (SEQ ID NO:9); d-DITWDQLWDLMK (SEQ ID NO:3);Dd-ITWDQLWDLMK (SEQ ID NO:3); ITWDQLWDLMK (amino acids 2-12 of SEQ IDNO:3); DITNal(1)DQLWDLMK-NH₂ (SEQ ID NO:118); DITWDQLNal(1)DLMK-NH₂ (SEQID NO:119); DITNal(2)DQLWDLMK-NH₂ (SEQ ID NO:120); DITWDQLNal(2)DLMK-NH₂(SEQ ID NO:121); DITChaDQLWDLMK-NH₂ (SEQ ID NO:122); DITWDQLChaDLMK-NH₂(SEQ ID NO:123); DITWDQLWDLM(OCH₃)K-NH₂ (SEQ ID NO:124);DITWDQLWDLM(SOCH₃)K-NH₂ (SEQ ID NO:125); DITWDQLWDLM(SO₂ CH₃)K-NH₂ (SEQID NO:126); DITWDQLW-Aib-LMK-NH₂ (SEQ ID NO:127); DITWDQLWDLMK-OH (SEQID NO:128); DITW-Aib-QLWDLMK-NH₂ (SEQ ID NO:130); DITW-Aib-QLWDLMK-OH(SEQ ID NO:132); DITWDQLWDLAK-OH (SEQ ID NO:143); DITWAQLWDLMK-NH₂ (SEQID NO:156); Ac-DITWDQLWDLMK-OH (SEQ ID NO:3); Succinimide-ITWDQLWDLMK-OH(amino acids 2-12 of SEQ ID NO:3); Cbz-ITWDQLWDLMK-OH (amino acids 2-12of SEQ ID NO:3); Succ-ITWDQLWDLMK-NH₂ (SEQ ID NO:159); andAc-DITWDQLWDLMK-NH₂ (SEQ ID NO:160).